Team:Groningen/Notebook/14 July 2009

From 2009.igem.org

[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]

Wet

GVP Cluster

www.sigmaaldrich.com

Plasmid Purification

Plasmid isolation was performed on the cultures of GVP and Terminator containing cells with the "Sygma-Aldrich™ [http://www.sigmaaldrich.com/life-science/molecular-biology/dna-and-rna-purification/plasmid-miniprep-kit.html GenElute™] Plasmid Miniprep Kit".

  • From each tube 5mL of culture was collected in a 2mL cup, and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded.
  • Cells were resuspended in 200μL Resuspension Solution by up and down pipetting.
  • To the mixture 200μL Lysis Solution was added, mixed by inverting the cup, and stored at room temperature for 5 minutes.
  • 350μL of Neutralisation Solution was added and the tubes inverted.
  • Cell debri was pelleted by centrifugation at full speed for 10 min.
  • Column was prepared by adding 500μL Column Preparation Solution and centrifuging for 1 min. full speed
  • The lysate was transfered to the column and centrifuged for 1 min. full speed
  • Column was washed with both washing solutions
  • Plasmids were eluted with 30μL MQ and stored in the fridge

Concentration of Plasmids

The concentration of isolated plasmid was determined with the use of a nano-drop.

[http://partsregistry.org/wiki/index.php/Part:BBa_I750016 GVP] eluted in MQ

  • 329.7 ng/μL
  • 1.84 (260/280)
  • 2.13 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 BBa_J23109] eluted in MQ

  • 174.9 ng/μL
  • 1.88 (260/280)
  • 1.92 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] eluted in MQ

  • 155.3 ng/μL
  • 1.85 (260/280)
  • 1.79 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 BBa_J23106] eluted in MQ

  • 219.4 ng/μL
  • 1.86 (260/280)
  • 1.93 (260/230)

Restriction of GVP and J23100/106/109 for assembly

A restriction was performed on the parts [http://partsregistry.org/wiki/index.php/Part:BBa_I750016 GVP], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 BBa_J23109], [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 BBa_J23106] as a first step in the construction of a new part containing the promoter and the GVP gene cluster.

Restriction mixture for GVP (fragment sizes: 6096 (GVP cluster), 3513)

  • 10 μL GVP plasmid (88.2 ng/μL)
  • 6μL MQ
  • 2μL Fast digest buffer
  • 1μL PstI fast digest enzyme
  • 1μL XbaI fast digest enzyme

Restriction mixture for J23109/100 (fragment sizes: 2096 (promoter + plasmid backbone), 887)

  • 10 μL promoter plasmid (61.4 ng/μL, 96.1 ng/μL)
  • 6μL MQ
  • 2μL Fast digest buffer
  • 1μL PstI fast digest enzyme
  • 1μL SpeI fast digest enzyme

- for J23106

  • 8 μL promoter plasmid (55.4 ng/μL)
  • 8μL MQ

Incubation for 30 min at 37°C.

The digests were run on a gel (1% agarose, TBE) and the bands containing the necessary fragments (6096, 2096) were excised.

Purification of fragments from agarose gel

Purification is done with the High pure PCR product purification kit (Roche).

  • 300 μL Binding buffer added for every 100 mg agarose
  • vortexed 30 s
  • 10 min incubation in 70°C (every 3 min vortexed 10 s)
  • 150 μL isopropanol added forevery 100 mg of agarose
  • vortexed 15 s
  • Contents pipetted into a filter tube
  • Centrifugation 60 s max speed (14600 rpm), flowthrough is discarded
  • 500 μL Wash buffer added, centrifugation 60 s max speed, flowthrough is discarded
  • 200 μL Wash buffer added, centrifugation 60 s max speed, flowthrough is discarded
  • 50 μL Elution buffer added, centrifugation 60 s max speed
  • Flowthrough contains the product DNA

Concentrations of DNA

Determined with a Nanodrop.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23109 BBa_J23109] eluted in Elution buffer

  • 3.5 ng/μL
  • 1.90 (260/280)
  • 0.34 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 BBa_J23100] eluted in Elution buffer

  • 4.6 ng/μL
  • 2.64 (260/280)
  • 0.42 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23106 BBa_J23106] eluted in Elution buffer

  • 2.5 ng/μL
  • 2.10 (260/280)
  • 0.07 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_I750016 GVP (1)] eluted in Elution buffer

  • 3.5 ng/μL
  • 4.42 (260/280)
  • 0.05 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_I750016 GVP (2)] eluted in Elution buffer

  • 5.2 ng/μL
  • 2.26 (260/280)
  • 0.07 (260/230)

[http://partsregistry.org/wiki/index.php?title=Part:BBa_I750016 GVP (3)] eluted in Elution buffer

  • 5.9 ng/μL
  • 2.36 (260/280)
  • 0.28 (260/230)

MQ was measured at the end: 1.8 ng/μL

Transporters

3nd PCR HmtA_Fw/HmtA_Rev We have no products on our gel but we do have ON cultures, we picked 3 for isolation.

Today we have the other primers as well. Since we are almost out of template DNA we preform colony PCR on the Colonies of the HmtA plate 3. With a dynazyme master mix (2 samples picked) and homebrew (2 samples picked). To see which ON culture to isolate plasmide from. and continue Cloning by mutating with mutation primer on the PCR product.

MasterMix3
Component Mg/K
MQ 28.5 uL
F mut1 2 uL
R mut2 2 uL
dNTP 2 uL
MgCl 3 uL
taq buffer KCl 5 uL
taq buffer (NH)SO 5 uL
taq polymerase 2.5 uL
PCR 3 program Temperature Time
Denaturing 95° 2.00 min
Start Cycles 25X
Denaturing 95° 30 sec
Annealing 55° 20 sec
Elongation 72° 2.10 min
End cycles
Final elongation 72° 10 min
Hold Forever

Vectors

Innoculated reaction tubes filled with 3 ml Ty or LB for overnight shaking 37°

  • pSB3K3 3ml kan LB and Ty
  • pSB1AC3 3ml amp LB and Ty
  • BBa_J23100 3ml amp LB and Ty
  • BBa_J23106 3ml amp LB and Ty
  • BBa_J23109 3ml amp LB and Ty


Dry

The arsenic accumulation model was worked out to the point where it supports pretty much all the kinds of operators we're interested in (at least the equilibrium model), and slightly more realistic parameters were used in the Simbiology model. The equilibrium calculator on the Wiki uses [http://en.wikipedia.org/wiki/Fixed_point_iteration fixed point iteration] to compute the equilibrium, so far this has proven to be quite fast and accurate (in addition it is full of sanity checks). And nicely enough the calculator on the Wiki so far agrees with the Simbiology model!


April
MTWTFSS
    [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 5]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/17_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 19]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_April_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]
May
MTWTFSS
        [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 3]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 6] [http://2009.igem.org/Team:Groningen/Notebook/7_May_2009 7] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 10]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/17_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 17]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 24]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 27] [http://2009.igem.org/Team:Groningen/Notebook/28_May_2009 28] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/31_May_2009&preload=Team:Groningen/Notebook/Day&action=edit 31]
June
MTWTFSS
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 4] [http://2009.igem.org/Team:Groningen/Notebook/5_June_2009 5] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 7]
[http://2009.igem.org/Team:Groningen/Notebook/8_June_2009 8] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 14]
[http://2009.igem.org/Team:Groningen/Notebook/15_June_2009 15] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 16] [http://2009.igem.org/Team:Groningen/Notebook/17_June_2009 17] [http://2009.igem.org/Team:Groningen/Notebook/18_June_2009 18] [http://2009.igem.org/Team:Groningen/Notebook/19_June_2009 19] [http://2009.igem.org/Team:Groningen/Notebook/20_June_2009 20] [http://2009.igem.org/Team:Groningen/Notebook/21_June_2009 21]
[http://2009.igem.org/Team:Groningen/Notebook/22_June_2009 22] [http://2009.igem.org/Team:Groningen/Notebook/23_June_2009 23] [http://2009.igem.org/Team:Groningen/Notebook/24_June_2009 24] [http://2009.igem.org/Team:Groningen/Notebook/25_June_2009 25] [http://2009.igem.org/Team:Groningen/Notebook/26_June_2009 26] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 28]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_June_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]
July
MTWTFSS
    [http://2009.igem.org/Team:Groningen/Notebook/1_July_2009 1] [http://2009.igem.org/Team:Groningen/Notebook/2_July_2009 2] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 5]
[http://2009.igem.org/Team:Groningen/Notebook/6_July_2009 6] [http://2009.igem.org/Team:Groningen/Notebook/7_July_2009 7] [http://2009.igem.org/Team:Groningen/Notebook/8_July_2009 8] [http://2009.igem.org/Team:Groningen/Notebook/9_July_2009 9] [http://2009.igem.org/Team:Groningen/Notebook/10_July_2009 10] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 12]
[http://2009.igem.org/Team:Groningen/Notebook/13_July_2009 13] [http://2009.igem.org/Team:Groningen/Notebook/14_July_2009 14] [http://2009.igem.org/Team:Groningen/Notebook/15_July_2009 15] [http://2009.igem.org/Team:Groningen/Notebook/16_July_2009 16] [http://2009.igem.org/Team:Groningen/Notebook/17_July_2009 17] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 19]
[http://2009.igem.org/Team:Groningen/Notebook/20_July_2009 20] [http://2009.igem.org/Team:Groningen/Notebook/21_July_2009 21] [http://2009.igem.org/Team:Groningen/Notebook/22_July_2009 22] [http://2009.igem.org/Team:Groningen/Notebook/23_July_2009 23] [http://2009.igem.org/Team:Groningen/Notebook/24_July_2009 24] [http://2009.igem.org/Team:Groningen/Notebook/25_July_2009 25] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_July_2009&preload=Team:Groningen/Notebook/Day&action=edit 26]
[http://2009.igem.org/Team:Groningen/Notebook/27_July_2009 27] [http://2009.igem.org/Team:Groningen/Notebook/28_July_2009 28] [http://2009.igem.org/Team:Groningen/Notebook/29_July_2009 29] [http://2009.igem.org/Team:Groningen/Notebook/30_July_2009 30] [http://2009.igem.org/Team:Groningen/Notebook/31_July_2009 31]
August
MTWTFSS
          [http://2009.igem.org/Team:Groningen/Notebook/1_August_2009 1] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_August_2009&preload=Team:Groningen/Notebook/Day&action=edit 2]
[http://2009.igem.org/Team:Groningen/Notebook/3_August_2009 3] [http://2009.igem.org/Team:Groningen/Notebook/4_August_2009 4] [http://2009.igem.org/Team:Groningen/Notebook/5_August_2009 5] [http://2009.igem.org/Team:Groningen/Notebook/6_August_2009 6] [http://2009.igem.org/Team:Groningen/Notebook/7_August_2009 7] [http://2009.igem.org/Team:Groningen/Notebook/8_August_2009 8] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_August_2009&preload=Team:Groningen/Notebook/Day&action=edit 9]
[http://2009.igem.org/Team:Groningen/Notebook/10_August_2009 10] [http://2009.igem.org/Team:Groningen/Notebook/11_August_2009 11] [http://2009.igem.org/Team:Groningen/Notebook/12_August_2009 12] [http://2009.igem.org/Team:Groningen/Notebook/13_August_2009 13] [http://2009.igem.org/Team:Groningen/Notebook/14_August_2009 14] [http://2009.igem.org/Team:Groningen/Notebook/15_August_2009 15] [http://2009.igem.org/Team:Groningen/Notebook/16_August_2009 16]
[http://2009.igem.org/Team:Groningen/Notebook/17_August_2009 17] [http://2009.igem.org/Team:Groningen/Notebook/18_August_2009 18] [http://2009.igem.org/Team:Groningen/Notebook/19_August_2009 19] [http://2009.igem.org/Team:Groningen/Notebook/20_August_2009 20] [http://2009.igem.org/Team:Groningen/Notebook/21_August_2009 21] [http://2009.igem.org/Team:Groningen/Notebook/22_August_2009 22] [http://2009.igem.org/Team:Groningen/Notebook/23_August_2009 23]
[http://2009.igem.org/Team:Groningen/Notebook/24_August_2009 24] [http://2009.igem.org/Team:Groningen/Notebook/25_August_2009 25] [http://2009.igem.org/Team:Groningen/Notebook/26_August_2009 26] [http://2009.igem.org/Team:Groningen/Notebook/27_August_2009 27] [http://2009.igem.org/Team:Groningen/Notebook/28_August_2009 28] [http://2009.igem.org/Team:Groningen/Notebook/29_August_2009 29] [http://2009.igem.org/Team:Groningen/Notebook/30_August_2009 30]
[http://2009.igem.org/Team:Groningen/Notebook/31_August_2009 31]
September
MTWTFSS
  [http://2009.igem.org/Team:Groningen/Notebook/1_September_2009 1] [http://2009.igem.org/Team:Groningen/Notebook/2_September_2009 2] [http://2009.igem.org/Team:Groningen/Notebook/3_September_2009 3] [http://2009.igem.org/Team:Groningen/Notebook/4_September_2009 4] [http://2009.igem.org/Team:Groningen/Notebook/5_September_2009 5] [http://2009.igem.org/Team:Groningen/Notebook/6_September_2009 6]
[http://2009.igem.org/Team:Groningen/Notebook/7_September_2009 7] [http://2009.igem.org/Team:Groningen/Notebook/8_September_2009 8] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 9] [http://2009.igem.org/Team:Groningen/Notebook/10_September_2009 10] [http://2009.igem.org/Team:Groningen/Notebook/11_September_2009 11] [http://2009.igem.org/Team:Groningen/Notebook/12_September_2009 12] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 13]
[http://2009.igem.org/Team:Groningen/Notebook/14_September_2009 14] [http://2009.igem.org/Team:Groningen/Notebook/15_September_2009 15] [http://2009.igem.org/Team:Groningen/Notebook/16_September_2009 16] [http://2009.igem.org/Team:Groningen/Notebook/17_September_2009 17] [http://2009.igem.org/Team:Groningen/Notebook/18_September_2009 18] [http://2009.igem.org/Team:Groningen/Notebook/19_September_2009 19] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 20]
[http://2009.igem.org/Team:Groningen/Notebook/21_September_2009 21] [http://2009.igem.org/Team:Groningen/Notebook/22_September_2009 22] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 23] [http://2009.igem.org/Team:Groningen/Notebook/24_September_2009 24] [http://2009.igem.org/Team:Groningen/Notebook/25_September_2009 25] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_September_2009&preload=Team:Groningen/Notebook/Day&action=edit 27]
[http://2009.igem.org/Team:Groningen/Notebook/28_September_2009 28] [http://2009.igem.org/Team:Groningen/Notebook/29_September_2009 29] [http://2009.igem.org/Team:Groningen/Notebook/30_September_2009 30]
October
MTWTFSS
      [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 1] [http://2009.igem.org/Team:Groningen/Notebook/2_October_2009 2] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 4]
[http://2009.igem.org/Team:Groningen/Notebook/5_October_2009 5] [http://2009.igem.org/Team:Groningen/Notebook/6_October_2009 6] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 11]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/14_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/17_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 18]
[http://2009.igem.org/Team:Groningen/Notebook/19_October_2009 19] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 25]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/31_October_2009&preload=Team:Groningen/Notebook/Day&action=edit 31]
November
MTWTFSS
            [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/1_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 1]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/2_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/3_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/4_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/5_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/6_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/7_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/8_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 8]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/9_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/10_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/11_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/12_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/13_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 13] [http://2009.igem.org/Team:Groningen/Notebook/14_November_2009 14] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/15_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 15]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/16_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 16] [http://2009.igem.org/Team:Groningen/Notebook/17_November_2009 17] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/18_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/19_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/20_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/21_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/22_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 22]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/23_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/25_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/26_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/27_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/28_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/29_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 29]
[http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/30_November_2009&preload=Team:Groningen/Notebook/Day&action=edit 30]