http://2009.igem.org/wiki/index.php?title=Special:Contributions/Grushina&feed=atom&limit=50&target=Grushina&year=&month=2009.igem.org - User contributions [en]2024-03-28T21:38:24ZFrom 2009.igem.orgMediaWiki 1.16.5http://2009.igem.org/Team:BIOTEC_Dresden/Team_v2Team:BIOTEC Dresden/Team v22010-05-23T17:29:07Z<p>Grushina: </p>
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<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
<br />
=== Our Team ===<br />
[[Image:Reduced_2IGEM_group2.jpg|600px|TEAM BIOTEC]]<br />
<br />
<br style="clear: both" /><br />
=== Who we are ===<br />
<br />
<div id="personsheader"><br />
'''Instructors'''<br />
<div id="persons"> [[Image:Petra.jpg|150px|thumb|[http://www.biotec.tu-dresden.de/cms/index.php?id=55: Petra Schwille] ]] </div><br />
<div id="persons"> [[Image:Francis.jpg|140px|thumb|[http://www.biotec.tu-dresden.de/stewart Francis Stewart]]] </div><br />
<br />
</div><br />
<br />
<br />
<div id="personsheader"><br />
'''Advisors'''<br />
<div id="persons"> [[Image:Kaj.jpg|150px|thumb|[http://www.linkedin.com/pub/kaj-bernhardt/8/964/531 Kaj Bernhardt]]] </div><br />
<div id="persons"> [[Image:yla.png|120px|thumb|[http://www.biotec.tu-dresden.de/cms/index.php?id=173 Ilaria Visco]]] </div><br />
<div id="persons"> [[Image:eugene.jpg|150px|thumb|Eugene Petrov]] </div><br />
<div id="persons"> [[Image:Salvatore.jpg|150px|thumb|[http://www.linkedin.com/pub/salvatore-loguercio/b/417/baa Salvatore Loguercio] ]] </div><br />
<div id="persons"> [[Image:Loic1.jpg|120px|thumb|Loic Royer]] </div><br />
</div><br />
<br />
<br />
<br />
<div id="personsheader"><br />
'''Students'''<br />
<div id="persons"> [[Image:Divya.jpg|150px|thumb|Divya Ail ]] </div><br />
<div id="persons"> [[Image:Tomek.png|140px|thumb|Tomek Sadowski ]] </div><br />
<div id="persons"> [[Image:Stanleypic.JPG|115px|thumb|Stanley Dinesh ]] </div><br />
<div id="persons"> [[Image:Prisha.jpg|130px|thumb|Priyanka Sharma]] </div><br />
</div><br />
<div id=personsheader"><br />
<div id="persons"> [[Image:Arnab.jpg|130px|thumb|Arnab Sen]] </div><br />
<div id="persons"> [[Image:Deepika.jpg|130px|thumb|Deepikaa Menon]] </div><br />
<div id="persons"> [[Image:Anja Grushina.JPG|140px|thumb|Anja Grushina]] </div><br />
<div id="persons"> [[Image:Dherde.jpg|140px|thumb|[http://dher.de Daniel Herde] ]] </div><br />
</div><br />
<br />
<br />
<br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/File:DSC_0842.JPGFile:DSC 0842.JPG2009-11-29T14:07:07Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Methods_VesiclesTeam:BIOTEC Dresden/Methods Vesicles2009-10-21T20:35:42Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Vesicles - Methods ===<br />
<br />
<br />
==== Setup of the microfluidic system ====<br />
<br />
<br />
The microfluidic system consists of a flow chamber made of Polydimethylsiloxane (PDMS) and a pump system that controls the flow rates of the various liquids into the chamber. Droplets are created within a defined space in the chamber and are propagated along a grid that allows containment and imaging. Two types of chambers have been used, differing in the geometry of the space where droplets were produced. One featured T-junction, and the other a V-junction (Fig1).<br />
<br />
<br />
Fig1: Different geometries of the intersection between aqueous and oil phase in flow chambers<br />
{|<br />
| [[Image:tchamber2.jpg|thumb|400|alt=t shaped junction|T shaped junction]]<br />
| [[Image:Vchamber.jpg|thumb|400|alt=v shaped junction|V shaped junction]]<br />
|}<br />
<br />
[[Image:Silicon wafer.jpg|200px|thumb|right|Silicon wafer we used for microfluidics chamber preparation]]<br />
Production of flow chambers:<br />
* mix PDMS and curing agent in 10:1 ratio<br />
* degas and pour on wafer with etched microstructures<br />
* polymerize on heat plate at 150 ºC for 30 min<br />
* add unpolymerized PDMS mixture to points on microstructure where microtube inlets are to be pierced<br />
* polymerize on heat plate at 150 ºC for 30 minutes<br />
* remove polymerized PDMS from wafer, cut to fit onto glass cover slide (24 x 60 mm), and use clean needles (0.8 mm) or laser cutter (Trotec Speedy 100TM) to pierce tube inlets<br />
* ionize PDMS and glass slide in plasma chamber for 30 sec to make it reactive<br />
* align PDMS on glass slide and seal<br />
* seal irreversibly by heating on plate at 60ºC for 6 hours<br />
<br />
<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowScriptAccess" value="always"></param><embed src="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1" type="application/x-shockwave-flash" allowfullscreen="true" allowScriptAccess="always" width="425" height="344"></embed></object> <html> <br><br />
<br />
<br />
The pumping system (ceDOSYS SP-4) allows control of syringes filled with aqueous material and surfactant treated with mineral oil, respectively. The syringes access the chamber via the tubing inlets. Two inlets are used to pump in material in the aqueous phase; the remaining one is used for the oil phase. The flow rates of the syringes are controlled via a ceDOSYS user interface software.<br />
<br />
Control via pump system:<br />
* two syringes are loaded with 1ml each of material in the aqueous phase; during the first trial, distilled water<br />
* another is filled with a 1ml solution of 0.5% span 80 in oil<br />
* use flow rate on ceDOSYS interface to flood the chamber first with oil phase<br />
* gradually introduce aqueous phase and modify rates of both phases until the shear stress breaks the aqueous phase into droplets at the T- or V- junctions in the respective chambers</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Results_VesiclesTeam:BIOTEC Dresden/Results Vesicles2009-10-21T20:28:31Z<p>Grushina: </p>
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<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Lipid Vesicles ===<br />
<br />
'''First approach: Microjetting'''<br />
<br />
Our initial idea was to make GUVs shooting aqueous material through a lipid bilayer using a microjetting setup (principle similar to blowing soap bubbles). <br />
<br />
[[Image:Our_1st_GUVs.jpg|300px]]<br />
<br />
Unfortunately, this method turned to be not technically reliable and the GUVs produced were not stable time wise and had broad size distribution.<br />
<br />
<br />
<br />
'''Second approach: Mixing in Microfluidic Chamber'''<br />
<br />
A microfluidic chamber that created an intersection between aqueous and oil phases produced vesicles that were uniform in size and stable for up to 5 hours. The vesicles are stabilized by a surfactant, Span 80.<br />
<br />
<br />
In the figure below vesicles are led into a grid by funnel shaped structures in the flow chamber (top). <br />
[[Image:vesicle_1.jpg|500px|vesicles created in microfluid chamber]]<br />
<br />
The video shows first see the part of the chamber where aqueous and oil phases meet. Oil flows from left to right in the channel that appears colorless. Two channels that carry material in the aqueous phase meet (this area is not visible in the image) and lead downwards towards the oil channel. The vesicles created are lead towards a network of channels that break the vesicles into smaller vesicles and eventually into a grid that stores them.<br />
<br />
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<html><br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Methods_VesiclesTeam:BIOTEC Dresden/Methods Vesicles2009-10-21T20:27:18Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Vesicles - Methods ===<br />
<br />
<br />
==== Setup of the microfluidic system ====<br />
<br />
<br />
The microfluidic system consists of a flow chamber made of Polydimethylsiloxane (PDMS) and a pump system that controls the flow rates of the various liquids into the chamber. Droplets are created within a defined space in the chamber and are propagated along a grid that allows containment and imaging. Two types of chambers have been used, differing in the geometry of the space where droplets were produced. One featured T-junction, and the other a V-junction (Fig1).<br />
<br />
<br />
Fig1: Different geometries of the intersection between aqueous and oil phase in flow chambers<br />
{|<br />
| [[Image:tchamber2.jpg|thumb|400|alt=t shaped junction|T shaped junction]]<br />
| [[Image:Vchamber.jpg|thumb|400|alt=v shaped junction|V shaped junction]]<br />
|}<br />
<br />
[[Image:Silicon wafer.jpg|200px|thumb|right|Silicon wafer we used for microfluidics chamber preparation]]<br />
Production of flow chambers:<br />
* mix PDMS and curing agent in 10:1 ratio<br />
* degas and pour on wafer with etched microstructures<br />
* polymerize on heat plate at 150 ºC for 30 min<br />
* add unpolymerized PDMS mixture to points on microstructure where microtube inlets are to be pierced<br />
* polymerize on heat plate at 150 ºC for 30 minutes<br />
* remove polymerized PDMS from wafer, cut to fit onto glass cover slide (24 x 60 mm), and use clean needles (0.8 mm) or laser cutter to pierce tube inlets<br />
* ionize PDMS and glass slide in plasma chamber for 30 sec to make it reactive<br />
* align PDMS on glass slide and seal<br />
* seal irreversibly by heating on plate at 60ºC for 6 hours<br />
<br />
<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowScriptAccess" value="always"></param><embed src="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1" type="application/x-shockwave-flash" allowfullscreen="true" allowScriptAccess="always" width="425" height="344"></embed></object> <html> <br><br />
<br />
<br />
The pumping system (ceDOSYS SP-4) allows control of syringes filled with aqueous material and surfactant treated with mineral oil, respectively. The syringes access the chamber via the tubing inlets. Two inlets are used to pump in material in the aqueous phase; the remaining one is used for the oil phase. The flow rates of the syringes are controlled via a ceDOSYS user interface software.<br />
<br />
Control via pump system:<br />
* two syringes are loaded with 1ml each of material in the aqueous phase; during the first trial, distilled water<br />
* another is filled with a 1ml solution of 0.5% span 80 in oil<br />
* use flow rate on ceDOSYS interface to flood the chamber first with oil phase<br />
* gradually introduce aqueous phase and modify rates of both phases until the shear stress breaks the aqueous phase into droplets at the T- or V- junctions in the respective chambers</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Methods_VesiclesTeam:BIOTEC Dresden/Methods Vesicles2009-10-21T20:26:39Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Vesicles - Methods ===<br />
<br />
<br />
==== Setup of the microfluidic system ====<br />
<br />
<br />
The microfluidic system consists of a flow chamber made of Polydimethylsiloxane (PDMS) and a pump system that controls the flow rates of the various liquids into the chamber. Droplets are created within a defined space in the chamber and are propagated along a grid that allows containment and imaging. Two types of chambers have been used, differing in the geometry of the space where droplets were produced. One featured T-junction, and the other a V-junction (Fig1).<br />
<br />
<br />
Fig1: Different geometries of the intersection between aqueous and oil phase in flow chambers<br />
{|<br />
| [[Image:tchamber2.jpg|thumb|400|alt=t shaped junction|T shaped junction]]<br />
| [[Image:Vchamber.jpg|thumb|400|alt=v shaped junction|V shaped junction]]<br />
|}<br />
<br />
[[Image:Silicon wafer.jpg|200px|thumb|right|Silicon wafer we used for microfluidics chamber preparation]]<br />
Production of flow chambers:<br />
* mix PDMS and curing agent in 10:1 ratio<br />
* degas and pour on wafer with etched microstructures<br />
* polymerize on heat plate at 150 ºC for 30 min<br />
* add unpolymerized PDMS mixture to points on microstructure where microtube inlets are to be pierced<br />
* polymerize on heat plate at 150 ºC for 30 minutes<br />
* remove polymerized PDMS from wafer, cut to fit onto glass cover slide (24 x 60 mm), and use clean needles (0.8 mm) or laser cutter to pierce tube inlets<br />
* ionize PDMS and glass slide in plasma chamber for 30 sec to make it reactive<br />
* align PDMS on glass slide and seal<br />
* seal irreversibly by heating on plate at 60ºC for 6 hours<br />
<br />
<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowScriptAccess" value="always"></param><embed src="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1" type="application/x-shockwave-flash" allowfullscreen="true" allowScriptAccess="always" width="425" height="344"></embed></object> <html> <br><br />
<br />
<br />
The pumping system (ceDOSYS SP-4) allows control of syringes filled with aqueous material and surfactant treated with mineral oil, respectively. The syringes access the chamber via the tubing inlets. Two inlets are used to pump in material in the aqueous phase; the remaining one is used for the oil phase. The flow rates of the syringes are controlled via a ceDOSYS user interface software.<br />
<br />
Control via pump system:<br />
* two syringes are loaded with 1ml each of material in the aqueous phase; during the first trial, distilled water<br />
* another is filled with a 1ml solution of 0.5% span 80 in oil<br />
* use flow rate on ceDOSYS interface to flood the chamber first with oil phase<br />
* gradually introduce aqueous phase and modify rates of both phases until the shear stress breaks the aqueous phase into droplets at the T- or V- junctions in the respective chambers<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Methods_VesiclesTeam:BIOTEC Dresden/Methods Vesicles2009-10-21T20:25:16Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Vesicles - Methods ===<br />
<br />
<br />
==== Setup of the microfluidic system ====<br />
<br />
<br />
The microfluidic system consists of a flow chamber made of Polydimethylsiloxane (PDMS) and a pump system that controls the flow rates of the various liquids into the chamber. Droplets are created within a defined space in the chamber and are propagated along a grid that allows containment and imaging. Two types of chambers have been used, differing in the geometry of the space where droplets were produced. One featured T-junction, and the other a V-junction (Fig1).<br />
<br />
<br />
Fig1: Different geometries of the intersection between aqueous and oil phase in flow chambers<br />
{|<br />
| [[Image:tchamber2.jpg|thumb|400|alt=t shaped junction|T shaped junction]]<br />
| [[Image:Vchamber.jpg|thumb|400|alt=v shaped junction|V shaped junction]]<br />
|}<br />
<br />
[[Image:Silicon wafer.jpg|200px|thumb|right|Silicon wafer we used for microfluidics chamber preparation]]<br />
Production of flow chambers:<br />
* mix PDMS and curing agent in 10:1 ratio<br />
* degas and pour on wafer with etched microstructures<br />
* polymerize on heat plate at 150 ºC for 30 min<br />
* add unpolymerized PDMS mixture to points on microstructure where microtube inlets are to be pierced<br />
* polymerize on heat plate at 150 ºC for 30 minutes<br />
* remove polymerized PDMS from wafer, cut to fit onto glass cover slide (24 x 60 mm), and use clean needles (0.8 mm) or laser cutter to pierce tube inlets<br />
* ionize PDMS and glass slide in plasma chamber for 30 sec to make it reactive<br />
* align PDMS on glass slide and seal<br />
* seal irreversibly by heating on plate at 60ºC for 6 hours<br />
<br />
<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowScriptAccess" value="always"></param><embed src="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1" type="application/x-shockwave-flash" allowfullscreen="true" allowScriptAccess="always" width="425" height="344"></embed></object> <html> <b><br />
<br />
<br />
The pumping system (ceDOSYS SP-4) allows control of syringes filled with aqueous material and surfactant treated with mineral oil, respectively. The syringes access the chamber via the tubing inlets. Two inlets are used to pump in material in the aqueous phase; the remaining one is used for the oil phase. The flow rates of the syringes are controlled via a ceDOSYS user interface software.<br />
<br />
Control via pump system:<br />
* two syringes are loaded with 1ml each of material in the aqueous phase; during the first trial, distilled water<br />
* another is filled with a 1ml solution of 0.5% span 80 in oil<br />
* use flow rate on ceDOSYS interface to flood the chamber first with oil phase<br />
* gradually introduce aqueous phase and modify rates of both phases until the shear stress breaks the aqueous phase into droplets at the T- or V- junctions in the respective chambers<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/File:Silicon_wafer.jpgFile:Silicon wafer.jpg2009-10-21T20:20:50Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Modeling_v2Team:BIOTEC Dresden/Modeling v22009-10-21T20:19:52Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
<br />
=== Looping probability simulation ===<br />
<br />
Interpolation formula [Stewart et. al. 1999] allows to calculate the probability of two cites of DNA to meet by looping. <br />
<br />
[[Image:Looping_probability_form.png]] <br />
[[Image:Looping_probability.png|350px|thumb|right|Fig.1 Normalized looping probability of DNA vs number of base pairs between the cites of interest]]<br />
<br />
The key parameters of the model are the distance between cites (in number of base pairs), persistence length of DNA (in nm) and the proximity of two DNA ends necessary for looping to occur (nm). Probability is expressed as the local molar concentration of one site with respect to the other. On the Fig. 1 results of a simulation according to this formula are shown. Persistence length is considered to be 50 nm [Hagerman, 1988], proximity of two cites is 10 nm (black line), compared to 0 nm (red line). Probability is normalized with respect to the maximum value (0.1194 10^-6 M) reached at ~400 bp. Both lines are consistent with experiment and show no looping below the persistence length (~150 bp). Our simulation (performed with MATLAB) proves that the optimal distance for looping to occur is ~400 bp and the probability of two cites to meet decreases with growing distance between them. As can be seen from the picture, accounting for non-zero distance between to cites to form a loop, gives higher probability of looping to happen.<br />
<br />
<br />
<br />
=== Theory behind FLP-FRT recombination ===<br />
<br />
<br />
In genetics, '''FLP-FRT recombination is a site-directed recombination technology used to manipulate an organism's DNA under controlled conditions in vivo'''. It is analogous to Cre-Lox recombination. It involves the recombination of sequences between short Flippase Recognition Target (FRT) sites by the Flippase recombination enzyme (FLP or Flp) derived from the 2µ plasmid of the baker's yeast Saccharomyces cerevisiae.<br />
<br />
The 34bp long FRT site sequence is : 5'-GAAGTTCCTATTCtctagaaaGTATAGGAACTTC-3'.<br />
Flippase (flp) binds to the 13-bp 5'-GAAGTTCCTATTC-3' and to the reverse complement of 5'-GTATAGGAACTTC-3' (5'-GAAGTTCCTATAC-3').<br />
The FRT site is cleaved just before 5'-tctagaaa-3', the 8bp asymmetric core region, on the top strand and behind this sequence on the bottom strand.[1]<br />
<br />
Several variant FRT sites exist. Recombination can occur between two identical FRT sites but generally not between non-identical FRT sites.<br />
<br />
Many available constructs include the sequence<br />
5'-GAAGTTCCTATTCC-3'<br />
immediately upstream the FRT site<br />
(resulting in 5'-GAAGTTCCTATTCCGAAGTTCCTATTCtctagaaaGTATAGGAACTTC-3')<br />
but this sequence is dispensable for recombination.<br />
<br />
Because the recombination activity can be targeted to only one target organ, or a low level of recombination activity can be used to consistently alter the DNA of only a subset of cells, '''FLP-FRT can be used to construct genetic mosaics in multicellular organisms'''. Using this technology, the loss or alteration of a gene can be studied in one target organ of interest, even if experimental animals could not survive the loss of the gene in other organs. <br />
<br />
The effect of altering a gene can also be studied over time, by using an inducible promoter to trigger the recombination activity late in development - this prevents the alteration from affecting overall development of an organ, and allows single cells lacking the gene to be compared to normal neighboring cells in the same environment.<br />
<br />
<br />
A very similar study using eukaryotic DNA:<br />
http://www.ncbi.nlm.nih.gov/pubmed/10581237<br />
<br />
kinetic analysis of Flp activity - DNA binding and recombination models:<br />
http://www.ncbi.nlm.nih.gov/pubmed/9813124<br />
<br />
Thermostability of Flp recombinase (We are using the F70L variant because it is sufficiently slow to give a time course):<br />
http://www.ncbi.nlm.nih.gov/pubmed/8932381<br />
<br />
http://www.recombineering.net/img/A202_01.gif<br />
<br />
'' pCAGGS-FLPe-IRESpuro expression vector.''<br />
<br />
<br />
<br />
Unlike transcriptional regulation, this method gives true all-or-none induction due to covalent modification of DNA by Flp recombinase. Determining the transfer curve of inter-FRT site distance versus average recombination time allows the onset of gene expression to be predicted. We then apply this [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2009&group=BIOTEC_Dresden Flp reporter system] as a powerful PoPS measurement device.<br />
<br />
<br />
<br />
=== Recombination pathway of FLP: design of mathematical model ===<br />
<br />
The recombination pathway upon which the<br />
mathematical model for recombination was based is shown in Figure 1. The model describes an excision<br />
reaction on a linear DNA substrate. The steps<br />
of DNA binding are well characterised for FLP. The enzyme binds the inverted repeat target<br />
site first as a monomer, which is then joined by a<br />
second monomer to form a dimer Andrews et al., 1987; Hoess et al., 1984; Mack et al., 1992). In the<br />
model, we have assumed that the protein is<br />
monomeric in solution, based on the behaviour of<br />
FLP and Cre in sucrose density gradients under<br />
similar buffer conditions to those used in this<br />
study (Abremski & Hoess, 1984; Qian et al., 1990).<br />
<br />
[[Image:Kinetic_FLP_1.png]]<br />
<br />
<br />
'''Figure 1'''Steps in FLP and Cre<br />
excision recombination reaction.<br />
Inverted repeat target sites are<br />
shown as open arrows. Recombinase<br />
monomers are shown as filled<br />
circles. Each step in the reaction is<br />
reversible and has a forward and a<br />
backward rate, indicated by small<br />
arrows. The forward and backward<br />
rate constants for each step are<br />
indicated on the Figure beside the<br />
arrows (small kn or k-n). The equilibrium<br />
constant for the conversion<br />
of one complex to another is given<br />
by the quotient of the backward<br />
and forward rate constants. Equilibrium<br />
constants are indicated (Kn).<br />
Names of species used for mathematical<br />
modelling are shown<br />
beside each complex (Ringrose et al., 1998, reproduced with permission).<br />
<br />
<br />
In our scheme, the second-order association rate<br />
constants for the binding of the first and second<br />
monomers are named k1 and k2, respectively. This<br />
process must occur twice to occupy both target<br />
sites of the excision substrate. In the model, all<br />
DNA binding and dissociation steps are described<br />
by k1 and k2, and their corresponding first-order<br />
dissociation rate constants, k-1 and k-2 (Figure 1).<br />
A reduced model, describing the sequential binding<br />
of recombinase monomers to a single full site<br />
target comprising two half sites, in terms of the<br />
four parameters k1, k2, k-1 and k-2 is described by<br />
equations (7) to (10) (see below).<br><br />
After DNA binding, the next step in the pathway<br />
is synapsis. FLP synapsis occurs by random collision<br />
(Beatty et al., 1986). In Figure 1, synapsis of the fully occupied<br />
excision substrate is described by a single first-order<br />
rate constant, k3. The model only allows for<br />
intramolecular synapsis, although in reality intermolecular<br />
recombination can also occur. However,<br />
for recombination assays, standard experimental<br />
conditions have been used, under which the frequency of<br />
intermolecular recombination is negligible. The<br />
multiple steps of catalysis are well characterised<br />
for FLP (for reviews, see Stark et al., 1992;<br />
Jayaram, 1994; Sadowski, 1995), and are described<br />
in Figure 1 by a single pair of rate constants, k4<br />
and k-4, the forward and back rates of catalysis,<br />
respectively.<br><br />
At present, we do not have a means of accurately<br />
measuring the formation of the synaptic<br />
complex. For this reason, we have further simplified the model by combining the constants k3 and<br />
k4 to give an apparent constant, k34,<br />
describing the rate of conversion of the fully<br />
bound substrate (SM4) into the excised synaptic<br />
complex (IEP) (Figure 1). The back rate, k-34,<br />
describes the reverse process. The relationship<br />
between k34, k-34 and their components k3, k-3, k4<br />
and k-4 is given by:<br><br />
<br />
[[Image:Eq6_FLP.png]]<br />
<br />
In this scheme, the dissociation of the synapse is<br />
represented as a reversal of its assembly: dissociation<br />
gives rise first to two products, each of<br />
which is bound by a recombinase dimer (Figure 1).<br />
This process is described by the first-order rate<br />
constant, k5. The dissociation of recombinase from<br />
DNA is assumed to occur in a stepwise manner,<br />
and is described in the model by the rate constants<br />
k-1 and k-2 (Figure 1). This assumption is based on<br />
the most simple and logical pathway. The dissociation<br />
mechanism of FLP has not been<br />
studied extensively, and there is disagreement in<br />
the literature regarding the steps involved. In experiments with synthetic Holliday<br />
junctions, it has been reported that the resolution<br />
of such structures requires two (Dixon &<br />
Sadowski, 1993), three (Qian & Cox, 1995) or four<br />
molecules of FLP (Lee et al., 1996). Based on experiments<br />
using full recombination substrates, Waite<br />
& Cox (1995) proposed a mechanism for FLP dissociation<br />
in which one or more monomers leave<br />
the synapse after recombination whilst others<br />
remain bound for longer. In the absence of a consensus<br />
on the mechanism of dissociation, Ringrose et al. (1998) proposed<br />
the mechanism shown in Figure 1, and point<br />
out that other mechanisms could easily be incorporated<br />
into the model by modification of<br />
equations (11) to (24) (see below).<br><br />
<br />
The DNA binding rate constants k1, k-1, k2, k-2, and their corresponding equilibrium constants K1 and K2, have been directly measured using the gel mobility shift assay. If the DNA binding constants are known, this leaves two pairs of unknown rate constants: k34 and k-34, and k5 and k-5 (Figure 1). The mathematical model describes a general excision recombination reaction in which four protein monomers are required to reversibly recombine a single substrate, giving two products. The values of all rate constants, and of protein and substrate input can be varied to represent specific cases.<br />
<br />
<br />
<br />
=== Simulation of FLP recombination ===<br />
<br />
<br />
<br />
[[Image:Kinetic_FLP_2.png]]<br />
<br />
<br />
'''Figure 8'''. Simulation of FLP recombination. a) Substrate<br />
titration (see below). The<br />
input values for protein, substrate, k1, k-1, k2 and k-2 are <br />
taken from Ringrose et al., (1998). Simulated<br />
curves for total nM excision product<br />
at three minutes and 60 minutes<br />
were fitted to the experimental<br />
data for FLP by simultaneous optimisation<br />
of the rate constants k34, k-34,<br />
k5 and k-5. Continuous lines, simulated<br />
recombination at three and<br />
60 minutes; 25.6 nM FLP, 0.05 to<br />
10 nM substrate. Open circles,<br />
recombination observed at three<br />
minutes. Filled circles,<br />
recombination observed at 60 minutes. b)<br />
Time course. The optimised parameters in Ringrose et al.,(1998) were used to<br />
simulate time course curves for<br />
FLP. Continuous<br />
line, simulated recombination time<br />
course at 25.6 nM FLP, 0.4 nM substrate;<br />
filled circles, data from time<br />
course experiment at 25.6 nM FLP, 0.4 nM substrate. c) Protein titration. Recombination time course curves were simulated for FLP at various protein inputs, with 0.4 nM substrate input. The simulated recombination after 60 minutes is plotted against protein concentration. Continuous line, simulated recombination parameters as in Ringrose et al.,(1998). Open circles, data points from protein titration experiment.<br />
<br />
<br />
<br />
'''Mathematical modelling I: DNA binding'''<br />
<br />
Using the nomenclature of Figure 1, the binding of<br />
recombinase monomers (M) to a full site substrate (S) is<br />
described by the kinetic equations:<br />
<br />
[[Image:eq1_FLP.png]]<br />
<br />
k1 and k-1 are the association and dissociation rate constants,<br />
respectively, for the species SM. k2 and k-2 are the<br />
association and dissociation rate constants for the species<br />
SM2. For the DNA binding model the assumption is<br />
made that the on and off rates for each half site, a and b,<br />
are identical.<br />
The evolution of species M, S, SM and SM2 over time<br />
is described by the differential equations:<br />
<br />
<br />
[[Image:eq2_FLP.png]]<br />
<br />
[[Image:eq3_FLP.png]]<br />
<br />
''dt'' is the time interval in seconds. The terms in square<br />
brackets refer to molar concentrations at time, t.<br />
Equations (7) to (10) were implemented in a Fortran 77<br />
code, and solved by finite difference numerical integration.<br />
The rate constants k1, k-1, k2 and k-2 were determined<br />
by fitting the computed time courses of the SM and SM2<br />
concentrations to the experimentally observed values<br />
simultaneously. Starting values of k1, k-1, k2 and k-2<br />
were determined by choosing values which gave an<br />
approximate agreement with the experimental data.<br />
Using these starting values, the model of equations (7) to<br />
(10) was integrated in time using a semi-implicit Euler<br />
scheme (Press et al., 1992). The time courses were<br />
fitted to the experimental data by minimising an error<br />
function using the method of Powell (1965), a derivative-free<br />
version of the conjugate gradient method. The error<br />
function was constructed by summing the squared deviations<br />
of the experimental and computed SM and SM2<br />
concentrations. As each set of rate constants corresponds<br />
to a unique solution of equations (7) to (10), and hence a<br />
given value of the error function, optimal values of the<br />
rate constants correspond to optimal agreement between<br />
experimental and modelled time courses.<br />
<br />
<br />
'''Mathematical modelling II: the<br />
recombination reaction'''<br />
<br />
The proposed pathway of recombination is shown in<br />
Figure 1. The names of most species and rate constants<br />
used in the model are shown in the Figure. Additional<br />
species described by the model and not shown in<br />
Figure 1 are as follows. The distinction is made in the<br />
model between SM2A (equation (14)), which has a single<br />
monomer bound at each target site, and SM2B (equation<br />
(15)), which has two monomers bound at one site, leaving<br />
the other site unoccupied. Only SM2B is indicated in<br />
Figure 1. Either SM2A or SM2B can give rise to SM3<br />
(equation (16)), in which one site is bound by two monomers<br />
while the other is bound by only one monomer.<br />
Later species which are also not shown in Figure 1<br />
describe the stepwise dissociation of monomers from<br />
DNA. EPM2 and LPM2 (Figure 1), both with two monomers<br />
bound, can each release a single monomer to give<br />
rise to EPM and LPM respectively (equations (21) and<br />
(22)). The release of the last bound monomer from EPM<br />
and LPM gives rise to EP and LP (equations (23) and<br />
(24); Figure 1).<br />
Based on classical kinetic equations, the differential<br />
equations describing the rate of change of each species<br />
over time (in seconds) are as follows:<br />
<br />
<br />
[[Image:Eq4_FLP.png]]<br />
<br />
[[Image:eq5_FLP.png]]<br />
<br />
<br />
Equations (11) to (24) were implemented and solved<br />
as described above for the DNA binding model. To find<br />
values for the unknown rate constants k34, k-34, k5 and<br />
k-5, computed time courses were fitted to experimental<br />
data by varying the input values of unknown rate constants.<br />
The quantity taken to represent the total excision<br />
product, EPtot, is given by IEP + EPM2 + EPM + EP.<br />
This quantity is considered to describe the experimentally<br />
observed excision product because only the linear<br />
excision product, and not the circle, is labelled and quanti<br />
fied. In addition, because recombination reactions are<br />
terminated with proteinase K, the DNA seen as excision<br />
product includes the DNA within the excised synaptic<br />
complex (IEP in the model) and all subsequent bound<br />
and unbound forms (EPM2, EPM and EP).<br></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Team_v2Team:BIOTEC Dresden/Team v22009-10-21T18:24:19Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
<br />
=== Our Team ===<br />
[[Image:Reduced_2IGEM_group2.jpg|500px|TEAM BIOTEC]]<br />
<br />
<br style="clear: both" /><br />
=== Who we are ===<br />
<br />
<div id="personsheader"><br />
'''Instructors'''<br />
<div id="persons"> [[Image:Petra.jpg|150px|thumb|[http://www.biotec.tu-dresden.de/cms/index.php?id=55: Petra Schwille] ]] </div><br />
<div id="persons"> [[Image:Francis.jpg|140px|thumb|[http://www.biotec.tu-dresden.de/stewart Francis Stewart]]] </div><br />
<br />
</div><br />
<br />
<br />
<div id="personsheader"><br />
'''Advisors'''<br />
<div id="persons"> [[Image:Salvatore.jpg|150px|thumb|[http://www.linkedin.com/pub/salvatore-loguercio/b/417/baa Salvatore Loguercio] ]] </div><br />
<div id="persons"> [[Image:Reduced_2IGEM_group2.jpg|150px|thumb|[http://www.biotec.tu-dresden.de/cms/index.php?id=173 Ilaria Visco]]] </div><br />
<div id="persons"> [[Image:Reduced_2IGEM_group2.jpg|150px|thumb|[http://www.linkedin.com/pub/kaj-bernhardt/8/964/531 Kaj Bernhardt]]] </div><br />
<div id="persons"> [[Image:Reduced_2IGEM_group2.jpg|150px|thumb|Loic Royer]] </div><br />
</div><br />
<br />
<br />
<br />
<div id="personsheader"><br />
'''Students'''<br />
<div id="persons"> [[Image:Divya.jpg|150px|thumb|Divya Ail ]] </div><br />
<div id="persons"> [[Image:Tomek.png|140px|thumb|Tomek Sadovski ]] </div><br />
<div id="persons"> [[Image:Stanleypic.JPG|105px|thumb|Stanley Dinesh ]] </div><br />
<div id="persons"> [[Image:Prisha.jpg|120px|thumb|Priyanka Sharma ]] </div><br />
<div id="persons"> [[Image:Reduced_2IGEM_group2.jpg|150px|thumb|Arnab Sen ]] </div><br />
<div id="persons"> [[Image:Deepika.jpg|140px|thumb|Deepikaa Menon]] </div><br />
<div id="persons"> [[Image:Anja Grushina.JPG|150px|thumb|Anja Grushina]] </div><br />
<div id="persons"> [[Image:Dherde.jpg|150px|thumb|[http://dher.de Daniel Herde] ]] </div><br />
</div><br />
<br />
<br />
<br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/File:Anja_Grushina.JPGFile:Anja Grushina.JPG2009-10-21T18:23:09Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Methods_VesiclesTeam:BIOTEC Dresden/Methods Vesicles2009-10-21T14:32:30Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Vesicles - Methods ===<br />
<br />
<br />
==== Setup of the microfluidic system ====<br />
<br />
<br />
The microfluidic system consists of a flow chamber made of Polydimethylsiloxane (PDMS) and a pump system that controls the flow rates of the various liquids into the chamber. Droplets are created within a defined space in the chamber and are propagated along a grid that allows containment and imaging. Two types of chambers have been used, differing in the geometry of the space where droplets were produced. One featured T-junction, and the other a V-junction (Fig1).<br />
<br />
<br />
Fig1: Different geometries of the intersection between aqueous and oil phase in flow chambers<br />
{|<br />
| [[Image:tchamber2.jpg|thumb|400|alt=t shaped junction|T shaped junction]]<br />
| [[Image:Vchamber.jpg|thumb|400|alt=v shaped junction|V shaped junction]]<br />
|}<br />
<br />
<br />
Production of flow chambers:<br />
* mix PDMS and curing agent in 10:1 ratio<br />
* degas and pour on wafer with etched microstructures<br />
* polymerize on heat plate at 150 ºC for 30 min<br />
* add unpolymerized PDMS mixture to points on microstructure where microtube inlets are to be pierced<br />
* polymerize on heat plate at 150 ºC for 30 minutes<br />
* remove polymerized PDMS from wafer, cut to fit onto glass cover slide (24 x 60 mm), and use clean needles (0.8 mm) or laser cutter to pierce tube inlets<br />
* ionize PDMS and glass slide in plasma chamber for 30 sec to make it reactive<br />
* align PDMS on glass slide and seal<br />
* seal irreversibly by heating on plate at 60ºC for 6 hours<br />
<br />
<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowScriptAccess" value="always"></param><embed src="http://www.youtube.com/v/gWoev1RP9SU&color1=0xb1b1b1&color2=0xcfcfcf&hl=en&feature=player_embedded&fs=1" type="application/x-shockwave-flash" allowfullscreen="true" allowScriptAccess="always" width="425" height="344"></embed></object> <html><br />
<br />
<br />
The pumping system (ceDOSYS SP-4) allows control of syringes filled with aqueous material and surfactant treated with mineral oil, respectively. The syringes access the chamber via the tubing inlets. Two inlets are used to pump in material in the aqueous phase; the remaining one is used for the oil phase. The flow rates of the syringes are controlled via a ceDOSYS user interface software.<br />
<br />
Control via pump system:<br />
* two syringes are loaded with 1ml each of material in the aqueous phase; during the first trial, distilled water<br />
* another is filled with a 1ml solution of 0.5% span 80 in oil<br />
* use flow rate on ceDOSYS interface to flood the chamber first with oil phase<br />
* gradually introduce aqueous phase and modify rates of both phases until the shear stress breaks the aqueous phase into droplets at the T- or V- junctions in the respective chambers<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Results_VesiclesTeam:BIOTEC Dresden/Results Vesicles2009-10-21T13:14:37Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Lipid Vesicles ===<br />
<br />
'''First approach: Microjetting'''<br />
<br />
Our initial idea was to make GUVs shooting aqueous material through a lipid bilayer using a microjetting setup (principle similar to blowing soap bubbles). <br />
<br />
[[Image:Our_1st_GUVs.jpg|300px]]<br />
<br />
Unfortunately, this method turned to be not reliable techically and the GUVs produced were not stable time wise and had broad size distribution.<br />
<br />
<br />
<br />
'''Second approach: Mixing in Microfluidic Chamber'''<br />
<br />
A microfluidic chamber that created an intersection between aqueous and oil phases produced vesicles that were uniform in size and stable for up to 5 hours. The vesicles are stabilized by a surfactant, Span 80.<br />
<br />
<br />
In the figure below vesicles are led into a grid by funnel shaped structures in the flow chamber (top). <br />
[[Image:vesicle_1.jpg|500px|vesicles created in microfluid chamber]]<br />
<br />
The video shows first see the part of the chamber where aqueous and oil phases meet. Oil flows from left to right in the channel that appears colorless. Two channels that carry material in the aqueous phase meet (this area is not visible in the image) and lead downwards towards the oil channel. The vesicles created are lead towards a network of channels that break the vesicles into smaller vesicles and eventually into a grid that stores them.<br />
<br />
<html><br />
<object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/iq1ZRdI3eto&hl=en&fs=1&"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/iq1ZRdI3eto&hl=en&fs=1&" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object><br />
<html><br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Modeling_v2Team:BIOTEC Dresden/Modeling v22009-10-21T13:05:50Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
<br />
=== Looping probability simulation ===<br />
<br />
Interpolation formula [Stewart et. al. 1999] allows to calculate the probability of two cites of DNA to meet by looping. <br />
<br />
[[Image:Looping_probability_form.png]]<br />
<br />
The key parameters of the model are the distance between cites (in number of base pairs), persistence length of DNA (in nm) and the proximity of two DNA ends necessary for looping to occur (nm). Probability is expressed as the local molar concentration of one site with respect to the other. On the Fig. 1 results of a simulation according to this formula are shown. Persistence length is considered to be 50 nm [Hagerman, 1988], proximity of two cites is 10 nm (black line), compared to 0 nm (red line). Probability is normalized with respect to the maximum value (0.1194 10^-6 M) reached at ~400 bp. Both lines are consistent with experiment and show no looping below the persistence length (~150 bp). Our simulation (performed with MATLAB) proves that the optimal distance for looping to occur is ~400 bp and the probability of two cites to meet decreases with growing distance between them. As can be seen from the picture, accounting for non-zero distance between to cites to form a loop, gives higher probability of looping to happen.<br />
<br />
[[Image:Looping_probability.png|Fig.1 Normalized looping probability of DNA vs number of base pairs between the cites of interest]]<br />
<br />
<br />
=== Theory behind FLP-FRT recombination ===<br />
<br />
<br />
In genetics, '''FLP-FRT recombination is a site-directed recombination technology used to manipulate an organism's DNA under controlled conditions in vivo'''. It is analogous to Cre-Lox recombination. It involves the recombination of sequences between short Flippase Recognition Target (FRT) sites by the Flippase recombination enzyme (FLP or Flp) derived from the 2µ plasmid of the baker's yeast Saccharomyces cerevisiae.<br />
<br />
The 34bp long FRT site sequence is : 5'-GAAGTTCCTATTCtctagaaaGTATAGGAACTTC-3'.<br />
Flippase (flp) binds to the 13-bp 5'-GAAGTTCCTATTC-3' and to the reverse complement of 5'-GTATAGGAACTTC-3' (5'-GAAGTTCCTATAC-3').<br />
The FRT site is cleaved just before 5'-tctagaaa-3', the 8bp asymmetric core region, on the top strand and behind this sequence on the bottom strand.[1]<br />
<br />
Several variant FRT sites exist. Recombination can occur between two identical FRT sites but generally not between non-identical FRT sites.<br />
<br />
Many available constructs include the sequence<br />
5'-GAAGTTCCTATTCC-3'<br />
immediately upstream the FRT site<br />
(resulting in 5'-GAAGTTCCTATTCCGAAGTTCCTATTCtctagaaaGTATAGGAACTTC-3')<br />
but this sequence is dispensable for recombination.<br />
<br />
Because the recombination activity can be targeted to only one target organ, or a low level of recombination activity can be used to consistently alter the DNA of only a subset of cells, '''FLP-FRT can be used to construct genetic mosaics in multicellular organisms'''. Using this technology, the loss or alteration of a gene can be studied in one target organ of interest, even if experimental animals could not survive the loss of the gene in other organs. <br />
<br />
The effect of altering a gene can also be studied over time, by using an inducible promoter to trigger the recombination activity late in development - this prevents the alteration from affecting overall development of an organ, and allows single cells lacking the gene to be compared to normal neighboring cells in the same environment.<br />
<br />
<br />
A very similar study using eukaryotic DNA:<br />
http://www.ncbi.nlm.nih.gov/pubmed/10581237<br />
<br />
kinetic analysis of Flp activity - DNA binding and recombination models:<br />
http://www.ncbi.nlm.nih.gov/pubmed/9813124<br />
<br />
Thermostability of Flp recombinase (We are using the F70L variant because it is sufficiently slow to give a time course):<br />
http://www.ncbi.nlm.nih.gov/pubmed/8932381<br />
<br />
http://www.recombineering.net/img/A202_01.gif<br />
<br />
'' pCAGGS-FLPe-IRESpuro expression vector.''<br />
<br />
<br />
<br />
Unlike transcriptional regulation, this method gives true all-or-none induction due to covalent modification of DNA by Flp recombinase. Determining the transfer curve of inter-FRT site distance versus average recombination time allows the onset of gene expression to be predicted. We then apply this [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2009&group=BIOTEC_Dresden Flp reporter system] as a powerful PoPS measurement device.<br />
<br />
<br />
<br />
=== Recombination pathway of FLP: design of mathematical model ===<br />
<br />
The recombination pathway upon which the<br />
mathematical model for recombination was based is shown in Figure 1. The model describes an excision<br />
reaction on a linear DNA substrate. The steps<br />
of DNA binding are well characterised for FLP. The enzyme binds the inverted repeat target<br />
site first as a monomer, which is then joined by a<br />
second monomer to form a dimer Andrews et al., 1987; Hoess et al., 1984; Mack et al., 1992). In the<br />
model, we have assumed that the protein is<br />
monomeric in solution, based on the behaviour of<br />
FLP and Cre in sucrose density gradients under<br />
similar buffer conditions to those used in this<br />
study (Abremski & Hoess, 1984; Qian et al., 1990).<br />
<br />
[[Image:Kinetic_FLP_1.png]]<br />
<br />
<br />
'''Figure 1'''Steps in FLP and Cre<br />
excision recombination reaction.<br />
Inverted repeat target sites are<br />
shown as open arrows. Recombinase<br />
monomers are shown as filled<br />
circles. Each step in the reaction is<br />
reversible and has a forward and a<br />
backward rate, indicated by small<br />
arrows. The forward and backward<br />
rate constants for each step are<br />
indicated on the Figure beside the<br />
arrows (small kn or k-n). The equilibrium<br />
constant for the conversion<br />
of one complex to another is given<br />
by the quotient of the backward<br />
and forward rate constants. Equilibrium<br />
constants are indicated (Kn).<br />
Names of species used for mathematical<br />
modelling are shown<br />
beside each complex (Ringrose et al., 1998, reproduced with permission).<br />
<br />
<br />
In our scheme, the second-order association rate<br />
constants for the binding of the first and second<br />
monomers are named k1 and k2, respectively. This<br />
process must occur twice to occupy both target<br />
sites of the excision substrate. In the model, all<br />
DNA binding and dissociation steps are described<br />
by k1 and k2, and their corresponding first-order<br />
dissociation rate constants, k-1 and k-2 (Figure 1).<br />
A reduced model, describing the sequential binding<br />
of recombinase monomers to a single full site<br />
target comprising two half sites, in terms of the<br />
four parameters k1, k2, k-1 and k-2 is described by<br />
equations (7) to (10) (see below).<br><br />
After DNA binding, the next step in the pathway<br />
is synapsis. FLP synapsis occurs by random collision<br />
(Beatty et al., 1986). In Figure 1, synapsis of the fully occupied<br />
excision substrate is described by a single first-order<br />
rate constant, k3. The model only allows for<br />
intramolecular synapsis, although in reality intermolecular<br />
recombination can also occur. However,<br />
for recombination assays, standard experimental<br />
conditions have been used, under which the frequency of<br />
intermolecular recombination is negligible. The<br />
multiple steps of catalysis are well characterised<br />
for FLP (for reviews, see Stark et al., 1992;<br />
Jayaram, 1994; Sadowski, 1995), and are described<br />
in Figure 1 by a single pair of rate constants, k4<br />
and k-4, the forward and back rates of catalysis,<br />
respectively.<br><br />
At present, we do not have a means of accurately<br />
measuring the formation of the synaptic<br />
complex. For this reason, we have further simplified the model by combining the constants k3 and<br />
k4 to give an apparent constant, k34,<br />
describing the rate of conversion of the fully<br />
bound substrate (SM4) into the excised synaptic<br />
complex (IEP) (Figure 1). The back rate, k-34,<br />
describes the reverse process. The relationship<br />
between k34, k-34 and their components k3, k-3, k4<br />
and k-4 is given by:<br><br />
<br />
[[Image:Eq6_FLP.png]]<br />
<br />
In this scheme, the dissociation of the synapse is<br />
represented as a reversal of its assembly: dissociation<br />
gives rise first to two products, each of<br />
which is bound by a recombinase dimer (Figure 1).<br />
This process is described by the first-order rate<br />
constant, k5. The dissociation of recombinase from<br />
DNA is assumed to occur in a stepwise manner,<br />
and is described in the model by the rate constants<br />
k-1 and k-2 (Figure 1). This assumption is based on<br />
the most simple and logical pathway. The dissociation<br />
mechanism of FLP has not been<br />
studied extensively, and there is disagreement in<br />
the literature regarding the steps involved. In experiments with synthetic Holliday<br />
junctions, it has been reported that the resolution<br />
of such structures requires two (Dixon &<br />
Sadowski, 1993), three (Qian & Cox, 1995) or four<br />
molecules of FLP (Lee et al., 1996). Based on experiments<br />
using full recombination substrates, Waite<br />
& Cox (1995) proposed a mechanism for FLP dissociation<br />
in which one or more monomers leave<br />
the synapse after recombination whilst others<br />
remain bound for longer. In the absence of a consensus<br />
on the mechanism of dissociation, Ringrose et al. (1998) proposed<br />
the mechanism shown in Figure 1, and point<br />
out that other mechanisms could easily be incorporated<br />
into the model by modification of<br />
equations (11) to (24) (see below).<br><br />
<br />
The DNA binding rate constants k1, k-1, k2, k-2, and their corresponding equilibrium constants K1 and K2, have been directly measured using the gel mobility shift assay. If the DNA binding constants are known, this leaves two pairs of unknown rate constants: k34 and k-34, and k5 and k-5 (Figure 1). The mathematical model describes a general excision recombination reaction in which four protein monomers are required to reversibly recombine a single substrate, giving two products. The values of all rate constants, and of protein and substrate input can be varied to represent specific cases.<br />
<br />
<br />
<br />
=== Simulation of FLP recombination ===<br />
<br />
<br />
<br />
[[Image:Kinetic_FLP_2.png]]<br />
<br />
<br />
'''Figure 8'''. Simulation of FLP recombination. a) Substrate<br />
titration (see below). The<br />
input values for protein, substrate, k1, k-1, k2 and k-2 are <br />
taken from Ringrose et al., (1998). Simulated<br />
curves for total nM excision product<br />
at three minutes and 60 minutes<br />
were fitted to the experimental<br />
data for FLP by simultaneous optimisation<br />
of the rate constants k34, k-34,<br />
k5 and k-5. Continuous lines, simulated<br />
recombination at three and<br />
60 minutes; 25.6 nM FLP, 0.05 to<br />
10 nM substrate. Open circles,<br />
recombination observed at three<br />
minutes. Filled circles,<br />
recombination observed at 60 minutes. b)<br />
Time course. The optimised parameters in Ringrose et al.,(1998) were used to<br />
simulate time course curves for<br />
FLP. Continuous<br />
line, simulated recombination time<br />
course at 25.6 nM FLP, 0.4 nM substrate;<br />
filled circles, data from time<br />
course experiment at 25.6 nM FLP, 0.4 nM substrate. c) Protein titration. Recombination time course curves were simulated for FLP at various protein inputs, with 0.4 nM substrate input. The simulated recombination after 60 minutes is plotted against protein concentration. Continuous line, simulated recombination parameters as in Ringrose et al.,(1998). Open circles, data points from protein titration experiment.<br />
<br />
<br />
<br />
'''Mathematical modelling I: DNA binding'''<br />
<br />
Using the nomenclature of Figure 1, the binding of<br />
recombinase monomers (M) to a full site substrate (S) is<br />
described by the kinetic equations:<br />
<br />
[[Image:eq1_FLP.png]]<br />
<br />
k1 and k-1 are the association and dissociation rate constants,<br />
respectively, for the species SM. k2 and k-2 are the<br />
association and dissociation rate constants for the species<br />
SM2. For the DNA binding model the assumption is<br />
made that the on and off rates for each half site, a and b,<br />
are identical.<br />
The evolution of species M, S, SM and SM2 over time<br />
is described by the differential equations:<br />
<br />
<br />
[[Image:eq2_FLP.png]]<br />
<br />
[[Image:eq3_FLP.png]]<br />
<br />
''dt'' is the time interval in seconds. The terms in square<br />
brackets refer to molar concentrations at time, t.<br />
Equations (7) to (10) were implemented in a Fortran 77<br />
code, and solved by finite difference numerical integration.<br />
The rate constants k1, k-1, k2 and k-2 were determined<br />
by fitting the computed time courses of the SM and SM2<br />
concentrations to the experimentally observed values<br />
simultaneously. Starting values of k1, k-1, k2 and k-2<br />
were determined by choosing values which gave an<br />
approximate agreement with the experimental data.<br />
Using these starting values, the model of equations (7) to<br />
(10) was integrated in time using a semi-implicit Euler<br />
scheme (Press et al., 1992). The time courses were<br />
fitted to the experimental data by minimising an error<br />
function using the method of Powell (1965), a derivative-free<br />
version of the conjugate gradient method. The error<br />
function was constructed by summing the squared deviations<br />
of the experimental and computed SM and SM2<br />
concentrations. As each set of rate constants corresponds<br />
to a unique solution of equations (7) to (10), and hence a<br />
given value of the error function, optimal values of the<br />
rate constants correspond to optimal agreement between<br />
experimental and modelled time courses.<br />
<br />
<br />
'''Mathematical modelling II: the<br />
recombination reaction'''<br />
<br />
The proposed pathway of recombination is shown in<br />
Figure 1. The names of most species and rate constants<br />
used in the model are shown in the Figure. Additional<br />
species described by the model and not shown in<br />
Figure 1 are as follows. The distinction is made in the<br />
model between SM2A (equation (14)), which has a single<br />
monomer bound at each target site, and SM2B (equation<br />
(15)), which has two monomers bound at one site, leaving<br />
the other site unoccupied. Only SM2B is indicated in<br />
Figure 1. Either SM2A or SM2B can give rise to SM3<br />
(equation (16)), in which one site is bound by two monomers<br />
while the other is bound by only one monomer.<br />
Later species which are also not shown in Figure 1<br />
describe the stepwise dissociation of monomers from<br />
DNA. EPM2 and LPM2 (Figure 1), both with two monomers<br />
bound, can each release a single monomer to give<br />
rise to EPM and LPM respectively (equations (21) and<br />
(22)). The release of the last bound monomer from EPM<br />
and LPM gives rise to EP and LP (equations (23) and<br />
(24); Figure 1).<br />
Based on classical kinetic equations, the differential<br />
equations describing the rate of change of each species<br />
over time (in seconds) are as follows:<br />
<br />
<br />
[[Image:Eq4_FLP.png]]<br />
<br />
[[Image:eq5_FLP.png]]<br />
<br />
<br />
Equations (11) to (24) were implemented and solved<br />
as described above for the DNA binding model. To find<br />
values for the unknown rate constants k34, k-34, k5 and<br />
k-5, computed time courses were fitted to experimental<br />
data by varying the input values of unknown rate constants.<br />
The quantity taken to represent the total excision<br />
product, EPtot, is given by IEP + EPM2 + EPM + EP.<br />
This quantity is considered to describe the experimentally<br />
observed excision product because only the linear<br />
excision product, and not the circle, is labelled and quanti<br />
fied. In addition, because recombination reactions are<br />
terminated with proteinase K, the DNA seen as excision<br />
product includes the DNA within the excised synaptic<br />
complex (IEP in the model) and all subsequent bound<br />
and unbound forms (EPM2, EPM and EP).<br></div>Grushinahttp://2009.igem.org/File:Looping_probability_form.pngFile:Looping probability form.png2009-10-21T12:53:43Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Looping_probability.pngFile:Looping probability.png2009-10-21T12:34:27Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/Notebook_VesiclesTeam:BIOTEC Dresden/Notebook Vesicles2009-10-13T17:23:50Z<p>Grushina: </p>
<hr />
<div>{{:Team:BIOTEC_Dresden/NewTemplate}}<br />
=== Gene Expression in GUVs ===<br />
<br />
The idea is pretty nice. So tell them here why they should care.<br />
<br />
<br />
<br />
'''[[Strategy_Vesicles|Strategy]]'''<br />
<br />
The proposed project plan.<br />
<br />
<br />
<br />
'''[[Methods_Vesicles|Methods]]'''<br />
<br />
Here the basic ideas are introduced.<br />
<br />
Microfluidics chambers<br />
<br />
In order to make vesicles we use a microfluidic chamber incorporated with pumping system, controlled by the software. The experiment is observed by means of Biozero fluorescent microscope.<br />
<br />
Microfluidic chamber<br />
<br />
The chamber consist of two parts: glass coverslide and a PDMS layer with the microstructure printed on it. Glass does not require any special preparation but plasma oven treatment for 30 seconds at 15 mBars. <br />
<br />
PDMS (Polydimethyloxane) is an organic, silicon based polymer. [SiO(CH3)2] is the monomer (Fig. 1). PDMS is chemically inert, transparent, stable at room temperature, non-toxic and non-flammable and thus can be used as a material for the chamber.<br />
<br />
<br />
In order to prepare PDMS layer one needs a silicon wafer with litographically etched microstructures of required configuration and PDMS mixed with silicon elastomer curing agent in proportion 10:1. For two wafers that we have (Fig.2) we used 12g of PDMS and 1.2g of curing agent. Those two substances have to be well-mixed, degassed and placed on the clean wafer. Polymerization takes about 30 minutes at 150C. In the end of heating drops of PDMS 2-3 mm high should be placed on top of the structure in the places where the inlets are supposed to be. The polymerized PDMS should be carefully removed from the wafer and the holes for the inlets should be made. There are two ways how one can do those holes: by means of 0.8 mm needle or using a laser cutter. Both methods have advantages and disadvantages. Using needle requires a lot of training and most of samples get cracks around the holes, which means that the chamber will leak. Laser cutting is more accurate but it's difficult to place the PDMS precisely. <br />
<br />
When PDMS layer with microstructure printed on it is ready one cuts out the needed size for a chamber and treats it in plasma oven for 30 seconds (together with cover glass). This treatment makes the surface hydrophilic. After that polymer layer is placed onto the glass and the chamber is baked at 60C for 6 hours. <br />
<br />
In order to check if the chamber is working one tests in under the light microscope with mineral oil. 1 mm tube connected with the syringe with oil is inserted in lower inlet and oil is to fill the whole chamber. When the camber is washed with oil one should check every upper inlet in the similar way. This allows to check if the chamber is not leaking at the inlets and if it is working properly. Also, this makes the surface more hydrophobic which facilitates vesicles formation.<br />
<br />
The next step is to connect the chamber with the pumping system. <br />
<br />
Pumping system<br />
<br />
Our setup allows simultaneous control of pumping rate of 4 syringes driven by the software.<br />
<br />
Measurement<br />
<br />
The chamber should be placed on the microscope sample holder, connected with the syringes (inlet 1 – oil, inlet 2 water and DNA plasmid, 3 – water and an expression kit). The microscope is focused on the spot where channels bringing oil and water meet. In order to make the vesicles one should play around with the flow rates. Usually the ratio of oil/water flow rate is about 1/0.75. The expression of GFP in the vesicles is proved by fluorescent microscopy.<br />
<br />
'''[[Results_Vesicles|Results]]'''<br />
<br />
Lab notes and obtained results.<br />
<br />
{{:Team:BIOTEC_Dresden/NewTemplateEnd}}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/TeamTeam:BIOTEC Dresden/Team2009-07-03T12:47:03Z<p>Grushina: </p>
<hr />
<div>== '''Who we are''' ==<br />
{|border = "0"<br />
|-<br />
|rowspan="3"|<br />
<br />
<br />
<br />
<br />
'''Instructors'''<br />
<br />
*Petra Schwille<br />
*Francis Stewart<br />
<br />
*Kaj Bernhardt<br />
<br />
'''Advisors:'''<br />
<br />
*Salvatore Loguercio<br />
*Ilaria Visco <br />
*Loic Royer<br />
<br />
'''Students:'''<br />
<br />
*Divya Ail <br />
*Tomek Sadovski <br />
*Stanley Dinesh <br />
*Priyanka Sharma <br />
*Arnab Sen <br />
*Deepikaa Menon<br />
*Anja Grushina<br />
*Daniel Herde<br />
*Carlos Coral<br />
<br />
<br />
|<br />
<gallery><br />
Image:Member1.jpg|Divya Ail<br />
Image:Member2.jpg|Tomek Sadovski<br />
Image:Member3.jpg|Stanley Dinesh<br />
Image:Member4.jpg|Priyanka Sharma<br />
Image:Member5.jpg|Arnab Sen<br />
Image:Member6.jpg|Deepikaa Menon<br />
Image:Member7.jpg|Anja Grushina<br />
Image:Member8.jpg|Daniel Herde<br />
Image:Member9.jpg|Carlos Coral<br />
</gallery><br />
|}<br />
<br />
== '''What we did''' ==<br />
<br />
<br />
<br />
<br />
== '''Where we're from''' ==<br />
[[Image:Mw_arms.jpg]]<br />
That's more or less it<br />
<br />
[[Image:DSC09167.JPG|300px]]<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/TeamTeam:BIOTEC Dresden/Team2009-07-03T12:46:39Z<p>Grushina: </p>
<hr />
<div>== '''Who we are''' ==<br />
{|border = "0"<br />
|-<br />
|rowspan="3"|<br />
<br />
<br />
<br />
<br />
'''Instructors'''<br />
<br />
*Petra Schwille<br />
*Francis Stewart<br />
<br />
*Kaj Bernhardt<br />
<br />
'''Advisors:'''<br />
<br />
*Salvatore Loguercio<br />
*Ilaria Visco <br />
*Loic Royer<br />
<br />
'''Students:'''<br />
<br />
*Divya Ail <br />
*Tomek Sadovski <br />
*Stanley Dinesh <br />
*Priyanka Sharma <br />
*Arnab Sen <br />
*Deepikaa Menon<br />
*Anja Grushina<br />
*Daniel Herde<br />
*Carlos Coral<br />
<br />
<br />
|<br />
<gallery><br />
Image:Member1.jpg|Divya Ail<br />
Image:Member2.jpg|Tomek Sadovski<br />
Image:Member3.jpg|Stanley Dinesh<br />
Image:Member4.jpg|Priyanka Sharma<br />
Image:Member5.jpg|Arnab Sen<br />
Image:Member6.jpg|Deepikaa Menon<br />
Image:Member7.jpg|Anja Grushina<br />
Image:Member8.jpg|Daniel Herde<br />
Image:Member9.jpg|Carlos Coral<br />
</gallery><br />
|}<br />
<br />
== '''What we did''' ==<br />
<br />
<br />
<br />
<br />
== '''Where we're from''' ==<br />
[[Image:Mw_arms.jpg]]<br />
That's more or less it<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}<br />
<br />
<br />
[[Image:DSC09167.JPG|300px]]</div>Grushinahttp://2009.igem.org/File:DSC09167.JPGFile:DSC09167.JPG2009-07-03T12:46:07Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/TeamTeam:BIOTEC Dresden/Team2009-06-15T12:09:38Z<p>Grushina: </p>
<hr />
<div>== '''Who we are''' ==<br />
{|border = "0"<br />
|-<br />
|rowspan="3"|<br />
<br />
<br />
<br />
<br />
'''Instructors'''<br />
<br />
*Petra Schwille<br />
*Francis Stewart<br />
<br />
*Kaj Bernhardt<br />
<br />
'''Advisors:'''<br />
<br />
*Salvatore Loguercio<br />
*Ilaria Visco <br />
*Loic Royer<br />
<br />
'''Students:'''<br />
<br />
*Divya Ail <br />
*Tomek Sadovski <br />
*Stanley Dinesh <br />
*Priyanka Sharma <br />
*Arnab Sen <br />
*Deepikaa Menon<br />
*Anja Grushina<br />
*Daniel Herde<br />
*Carlos Coral<br />
<br />
<br />
|<br />
<gallery><br />
Image:Member1.jpg|Divya Ail<br />
Image:Member2.jpg|Tomek Sadovski<br />
Image:Member3.jpg|Stanley Dinesh<br />
Image:Member4.jpg|Priyanka Sharma<br />
Image:Member5.jpg|Arnab Sen<br />
Image:Member6.jpg|Deepikaa Menon<br />
Image:Member7.jpg|Anja Grushina<br />
Image:Member8.jpg|Daniel Herde<br />
Image:Member9.jpg|Carlos Coral<br />
</gallery><br />
|}<br />
<br />
== '''What we did''' ==<br />
<br />
<br />
<br />
<br />
== '''Where we're from''' ==<br />
[[Image:Mw_arms.jpg]]<br />
That's more or less it<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/NotebookTeam:BIOTEC Dresden/Notebook2009-06-03T16:59:38Z<p>Grushina: </p>
<hr />
<div>== Group 1: Cloning ==<br />
<br />
<br />
<br />
== Group 2: Nucleation ==<br />
<br />
<br />
<br />
== Group 3: GUVs ==<br />
<br />
<br />
Our first GUVs that had enough luck to appear<br />
<br />
[[Image:Our_1st_GUVs.jpg]]<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/ModelingTeam:BIOTEC Dresden/Modeling2009-06-03T16:59:23Z<p>Grushina: </p>
<hr />
<div><!--- The Mission, Experiments ---><br />
<br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/PartsTeam:BIOTEC Dresden/Parts2009-06-03T16:59:10Z<p>Grushina: </p>
<hr />
<div><br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/ProjectTeam:BIOTEC Dresden/Project2009-06-03T16:58:50Z<p>Grushina: </p>
<hr />
<div>== '''Overall project''' ==<br />
<br />
Our project: Biologically synthesized silver tags in artificial cells <br />
<br />
== Project Details==<br />
<br />
<br />
<br />
<br />
=== Group 1: Cloning ===<br />
<br />
=== Group 2: Nucleation of silver ===<br />
<br />
=== Group 3: GUVs ===<br />
<br />
<br />
<br />
<br />
<br />
=== The Experiments ===<br />
<br />
<br />
<br />
== Results ==<br />
<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/TeamTeam:BIOTEC Dresden/Team2009-06-03T16:58:33Z<p>Grushina: </p>
<hr />
<div>== '''Who we are''' ==<br />
{|border = "0"<br />
|-<br />
|rowspan="3"|<br />
<br />
<br />
<br />
<br />
'''Instructors'''<br />
<br />
*Petra Schwille<br />
*Francis Stewart<br />
<br />
*Kaj Bernhardt<br />
<br />
'''Advisors:'''<br />
<br />
*Salvatore Loguercio<br />
*Ilaria Visco <br />
<br />
<br />
'''Students:'''<br />
<br />
*Divya Ail <br />
*Tomek Sadovski <br />
*Stanley Dinesh <br />
*Priyanka Sharma <br />
*Arnab Sen <br />
*Deepikaa Menon<br />
*Anja Grushina<br />
*Daniel Herde<br />
*Carlos Coral<br />
<br />
<br />
|<br />
<gallery><br />
Image:Member1.jpg|Divya Ail<br />
Image:Member2.jpg|Tomek Sadovski<br />
Image:Member3.jpg|Stanley Dinesh<br />
Image:Member4.jpg|Priyanka Sharma<br />
Image:Member5.jpg|Arnab Sen<br />
Image:Member6.jpg|Deepikaa Menon<br />
Image:Member7.jpg|Anja Grushina<br />
Image:Member8.jpg|Daniel Herde<br />
Image:Member9.jpg|Carlos Coral<br />
</gallery><br />
|}<br />
<br />
== '''What we did''' ==<br />
<br />
<br />
<br />
<br />
== '''Where we're from''' ==<br />
[[Image:Mw_arms.jpg]]<br />
That's more or less it<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_DresdenTeam:BIOTEC Dresden2009-06-03T16:58:14Z<p>Grushina: </p>
<hr />
<div>{|align="justify"<br />
<br />
|We are 9 people from Dresden who study Nanobiophysics, Molecular Engineering and Physics. We came from different countries: Germany, India, Russia, Poland and Colombia. Our backgrounds are biology, biotechnology and physics. We are cool - soon we'll provide some evidence with our pictures and the project details :)<br />
|[[Image:Biotec tudd logo rgb.jpg|50px|right|frame]]<br />
|-<br />
|<br />
Our project: Biologically synthesized silver tags in artificial cells<br />
<br />
<br />
|[[Image:Team_biotec.jpg|right|frame|http://particlezoo.net/]]<br />
|-<br />
|<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#808080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/NotebookTeam:BIOTEC Dresden/Notebook2009-06-03T16:57:21Z<p>Grushina: </p>
<hr />
<div>== Group 1: Cloning ==<br />
<br />
<br />
<br />
== Group 2: Nucleation ==<br />
<br />
<br />
<br />
== Group 3: GUVs ==<br />
<br />
<br />
Our first GUVs that had enough luck to appear<br />
<br />
[[Image:Our_1st_GUVs.jpg]]<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/NotebookTeam:BIOTEC Dresden/Notebook2009-06-03T16:56:37Z<p>Grushina: </p>
<hr />
<div>== Group 1: Cloning ==<br />
<br />
<br />
<br />
== Group 2: Nucleation ==<br />
<br />
<br />
<br />
== Group 3: GUVs ==<br />
<br />
<br />
Our first GUVs that had a luck to appear<br />
<br />
Image:Our_1st_GUVs.jpg<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/File:Our_1st_GUVs.jpgFile:Our 1st GUVs.jpg2009-06-03T16:52:36Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/NotebookTeam:BIOTEC Dresden/Notebook2009-06-03T16:51:15Z<p>Grushina: </p>
<hr />
<div>Group 1: Cloning<br />
<br />
Group 2: Nucleation<br />
<br />
Group 3: GUVs<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/ModelingTeam:BIOTEC Dresden/Modeling2009-06-03T16:51:04Z<p>Grushina: </p>
<hr />
<div><!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/PartsTeam:BIOTEC Dresden/Parts2009-06-03T16:50:50Z<p>Grushina: </p>
<hr />
<div><br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/ProjectTeam:BIOTEC Dresden/Project2009-06-03T16:50:16Z<p>Grushina: </p>
<hr />
<div>== '''Overall project''' ==<br />
<br />
Our project: Biologically synthesized silver tags in artificial cells <br />
<br />
== Project Details==<br />
<br />
<br />
<br />
<br />
=== Group 1: Cloning ===<br />
<br />
=== Group 2: Nucleation of silver ===<br />
<br />
=== Group 3: GUVs ===<br />
<br />
<br />
<br />
<br />
<br />
=== The Experiments ===<br />
<br />
<br />
<br />
== Results ==<br />
<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/TeamTeam:BIOTEC Dresden/Team2009-06-03T16:49:58Z<p>Grushina: </p>
<hr />
<div>== '''Who we are''' ==<br />
{|border = "0"<br />
|-<br />
|rowspan="3"|<br />
<br />
<br />
<br />
<br />
'''Instructors'''<br />
<br />
*Petra Schwille<br />
*Francis Stewart<br />
<br />
*Kaj Bernhardt<br />
<br />
'''Advisors:'''<br />
<br />
*Salvatore Loguercio<br />
*Ilaria Visco <br />
<br />
<br />
'''Students:'''<br />
<br />
*Divya Ail <br />
*Tomek Sadovski <br />
*Stanley Dinesh <br />
*Priyanka Sharma <br />
*Arnab Sen <br />
*Deepikaa Menon<br />
*Anja Grushina<br />
*Daniel Herde<br />
*Carlos Coral<br />
<br />
<br />
|<br />
<gallery><br />
Image:Member1.jpg|Divya Ail<br />
Image:Member2.jpg|Tomek Sadovski<br />
Image:Member3.jpg|Stanley Dinesh<br />
Image:Member4.jpg|Priyanka Sharma<br />
Image:Member5.jpg|Arnab Sen<br />
Image:Member6.jpg|Deepikaa Menon<br />
Image:Member7.jpg|Anja Grushina<br />
Image:Member8.jpg|Daniel Herde<br />
Image:Member9.jpg|Carlos Coral<br />
</gallery><br />
|}<br />
<br />
== '''What we did''' ==<br />
<br />
<br />
<br />
<br />
== '''Where we're from''' ==<br />
[[Image:Mw_arms.jpg]]<br />
That's more or less it<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_DresdenTeam:BIOTEC Dresden2009-06-03T16:48:11Z<p>Grushina: </p>
<hr />
<div>{|align="justify"<br />
<br />
|We are 9 people from Dresden who study Nanobiophysics, Molecular Engineering and Physics. We came from different countries: Germany, India, Russia, Poland and Colombia. Our backgrounds are biology, biotechnology and physics. We are cool - soon we'll provide some evidence with our pictures and the project details :)<br />
|[[Image:Biotec tudd logo rgb.jpg|50px|right|frame]]<br />
|-<br />
|<br />
Our project: Biologically synthesized silver tags in artificial cells<br />
<br />
<br />
|[[Image:Team_biotec.jpg|right|frame|http://particlezoo.net/]]<br />
|-<br />
|<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#000000;background-color:#008080;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/File:Biotec_tudd_logo_rgb.jpgFile:Biotec tudd logo rgb.jpg2009-06-03T16:39:07Z<p>Grushina: uploaded a new version of "Image:Biotec tudd logo rgb.jpg"</p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_DresdenTeam:BIOTEC Dresden2009-06-03T16:37:22Z<p>Grushina: </p>
<hr />
<div>{|align="justify"<br />
<br />
|We are 9 people from Dresden who study Nanobiophysics, Molecular Engineering and Physics. We came from different countries: Germany, India, Russia, Poland and Colombia. Our backgrounds are biology, biotechnology and physics. We are cool - soon we'll provide some evidence with our pictures and the project details :)<br />
|[[Image:Biotec tudd logo rgb.jpg|50px|right|frame]]<br />
|-<br />
|<br />
Our project: Biologically synthesized silver tags in artificial cells<br />
<br />
<br />
|[[Image:Team_biotec.jpg|right|frame|http://particlezoo.net/]]<br />
|-<br />
|<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_DresdenTeam:BIOTEC Dresden2009-06-03T16:31:36Z<p>Grushina: </p>
<hr />
<div>{|align="justify"<br />
<br />
|We are 9 people from Dresden who study Nanobiophysics, Molecular Engineering and Physics. We came from different countries: Germany, India, Russia, Poland and Colombia. Our backgrounds are biology, biotechnology and physics. We are cool - soon we'll provide some evidence with our pictures and the project details :)<br />
|[[Image:Biotec tudd logo rgb.jpg|200px|right|frame]]<br />
|-<br />
|<br />
Our project: Biologically synthesized silver tags in artificial cells<br />
<br />
<br />
|[[Image:Team_biotec.jpg|right|frame|http://particlezoo.net/]]<br />
|-<br />
|<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_DresdenTeam:BIOTEC Dresden2009-06-03T16:30:54Z<p>Grushina: </p>
<hr />
<div>{|align="justify"<br />
<br />
|We are 9 people from Dresden who study Nanobiophysics, Molecular Engineering and Physics. We came from different countries: Germany, India, Russia, Poland and Colombia. Our backgrounds are biology, biotechnology and physics. We are cool - soon we'll provide some evidence with our pictures and the project details :)<br />
|[[Image:Biotec tudd logo rgb.jpg|200px|right|frame]]<br />
|-<br />
|<br />
Our project: Biologically synthesized silver tags in artificial cells<br />
<br />
<br />
|[[Image:Team_biotec.jpg|right|frame|]]<br />
|-<br />
|<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/Team:BIOTEC_DresdenTeam:BIOTEC Dresden2009-06-03T12:07:03Z<p>Grushina: </p>
<hr />
<div>{|align="justify"<br />
<br />
|We are 9 people from Dresden who study Nanobiophysics, Molecular Engineering and Physics. We came from different countries: Germany, India, Russia, Poland and Colombia. Our backgrounds are biology, biotechnology and physics. We are cool - soon we'll provide some evidence with our pictures and the project details :)<br />
|[[Image:Biotec tudd logo rgb.jpg|200px|right|frame]]<br />
|-<br />
|<br />
Our project: Biologically synthesized silver tags in artificial cells<br />
<br />
<br />
|[[Image:Team_biotec.jpg|right|frame|]]<br />
|-<br />
|<br />
|align="center"|[[Team:BIOTEC_Dresden | Team Example]]<br />
|}<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/File:Team_biotec.jpgFile:Team biotec.jpg2009-06-03T12:06:24Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Member8.jpgFile:Member8.jpg2009-06-03T12:05:47Z<p>Grushina: uploaded a new version of "Image:Member8.jpg"</p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Member8.jpgFile:Member8.jpg2009-06-03T12:05:14Z<p>Grushina: uploaded a new version of "Image:Member8.jpg"</p>
<hr />
<div></div>Grushinahttp://2009.igem.org/Team:BIOTEC_Dresden/TeamTeam:BIOTEC Dresden/Team2009-06-03T12:03:31Z<p>Grushina: </p>
<hr />
<div>== '''Who we are''' ==<br />
{|border = "0"<br />
|-<br />
|rowspan="3"|<br />
<br />
<br />
<br />
<br />
'''Instructors'''<br />
<br />
*Petra Schwille<br />
*Francis Stewart<br />
<br />
*Kaj Bernhardt<br />
<br />
'''Advisors:'''<br />
<br />
*Salvatore Loguercio<br />
*Ilaria Visco <br />
<br />
<br />
'''Students:'''<br />
<br />
*Divya Ail <br />
*Tomek Sadovski <br />
*Stanley Dinesh <br />
*Priyanka Sharma <br />
*Arnab Sen <br />
*Deepikaa Menon<br />
*Anja Grushina<br />
*Daniel Herde<br />
*Carlos Coral<br />
<br />
<br />
|<br />
<gallery><br />
Image:Member1.jpg|Divya Ail<br />
Image:Member2.jpg|Tomek Sadovski<br />
Image:Member3.jpg|Stanley Dinesh<br />
Image:Member4.jpg|Priyanka Sharma<br />
Image:Member5.jpg|Arnab Sen<br />
Image:Member6.jpg|Deepikaa Menon<br />
Image:Member7.jpg|Anja Grushina<br />
Image:Member8.jpg|Daniel Herde<br />
Image:Member9.jpg|Carlos Coral<br />
</gallery><br />
|}<br />
<br />
== '''What we did''' ==<br />
<br />
<br />
<br />
<br />
== '''Where we're from''' ==<br />
[[Image:Mw_arms.jpg]]<br />
That's more or less it<br />
<br />
<!--- The Mission, Experiments ---><br />
<br />
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"<br />
!align="center"|[[Team:BIOTEC_Dresden|Home]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Team|The Team]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Project|The Project]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Parts|Parts Submitted to the Registry]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Modeling|Modeling]]<br />
!align="center"|[[Team:BIOTEC_Dresden/Notebook|Notebook]]<br />
|}</div>Grushinahttp://2009.igem.org/File:Member9.jpgFile:Member9.jpg2009-06-03T12:02:20Z<p>Grushina: uploaded a new version of "Image:Member9.jpg"</p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Member9.jpgFile:Member9.jpg2009-06-03T12:01:44Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Member8.jpgFile:Member8.jpg2009-06-03T12:01:17Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Member7.jpgFile:Member7.jpg2009-06-03T12:00:56Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Member6.jpgFile:Member6.jpg2009-06-03T12:00:31Z<p>Grushina: </p>
<hr />
<div></div>Grushinahttp://2009.igem.org/File:Member5.jpgFile:Member5.jpg2009-06-03T12:00:12Z<p>Grushina: </p>
<hr />
<div></div>Grushina