Team:Warsaw/Calendar-Main/13 July 2009

From 2009.igem.org

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===Cloning the p53 coding sequence===
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<html>
-
 
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'''Marcin'''
+
-
 
+
-
Task 1:
+
-
 
+
-
*Prepare PCR reaction to amplified p53 coding sequence.
+
-
Methods:
+
-
* DNA matrix dilution:
+
-
1 &mi;l of plasmid solution (isolated in 11.07.09) was taken and diluted with 99 mi&;l of MQ water
+
-
* PCR mixture composition:
+
-
# proper mixture 1: 0.25 &mu;l primer 1 (50 nM; Oligo.pl), 0.25 &mu;l primer 2 (50 nM; Oligo.pl), 1.5 &mu;l dNTPs (20 &mu;M ;Fermentas), 0.5 &mu;l Pfu turbo polymerase (KNGiE), 2.5 &mu;l Pfu Turbo Buffer (Fermentas), 2.5 &mu;l MgSO<sub>4</sub> (20 &mu;M; Fermentas), 1 &mu;l DNA matrix, 16.5 &mu;l MQ water
+
-
# proprer mixture 2: 0.25 &mu;l primer 1 (50 nM; Oligo.pl), 0.25 &mu;l primer 2 (50 nM; Oligo.pl), 1.5 &mu;l dNTPs (20 &mu;M ;Fermentas), 0.5 &mu;l Pfu turbo polymerase (KNGiE), 2.5 &mu;l Pfu Turbo Buffer (Fermentas), 2.5 &mu;l MgSO<sub>4</sub> (20 &mu;M; Fermentas), 2 &mu;l DNA matrix, 15.5 &mu;l MQ water
+
-
# Negative control: the same as proper mixture 1, the only distinction is lack of the DNA matrix.
+
-
* Program:
+
-
 
+
-
p53 (detailed destription is [https://2009.igem.org/Team:Warsaw/Calendar-Main/9_July_2009 here])
+
-
 
+
-
Results:
+
-
 
+
-
*Clean-up the PCR products
+
-
 
+
-
Procedure: DNA was purified using the A&A clean-up kit. Detailed procedure is described [http://www.aabiot.com/products/dna_purification/dna_fragments/clean_up/protocol_clean_up.pdf here]
+
-
After clean-up 1 &mu;l of purified PCR product was loaded into gel and photographed. Unfortunatelly reaction 1 (2 &mu;l of DNA matrix) was abortive - there is no product in the gel.
+
<h3><div style="text-align: center;">Cloning of p53 coding sequence</div></h3>
-
 
+
<h4>Marcin</h4>
-
[[image:PCR_reaction-p53_13_07_09.png|thumb|center|300px|verification of PCR reaction and clean-up procedure]]
+
<br/>
-
 
+
<p>Task 1:</p>
-
Task 2:
+
<ul>
-
 
+
<li>Prepare PCR reaction to amplified p53 coding sequence.</li>
-
*Restriction digest of p53 coding sequence.
+
</ul>
-
Methods:
+
<p>Methods:</p>
-
*Control digest using PvuII  
+
<ul>
-
** Reaction mixture composition: 1 &mu;l purified PCR product, 0.5 &mu;l PvuII (Fermentas), 2 &mu;l Buffer Green (Fermentas), 15.5 &mu;l MQ water
+
<li>DNA template dilution:</li>
-
*Digest for subsequent cloning using XbaI
+
</ul>
-
** Reaction mixture composition: 10 &mu;l purified PCR product, 1 &mu;l XbaI (Fermentas), 5 &mu;l Buffer Tango (Fermentas), 34.5 &mu;l MQ water
+
<p>1 &mi;l of plasmid solution (isolated in 11.07.09) was taken and diluted with 99 mi&;l of MQ water</p>
-
* Both reaction were perform in the same condition:
+
<ul>
-
Program:
+
<li>PCR mixture composition:</li>
-
 
+
<ol>
-
digest:
+
<li>proper mixture 1:
 +
<pre>0.25 &mu;l primer 1 (50 nM; Oligo.pl)
 +
0.25 &mu;l primer 2 (50 nM; Oligo.pl)
 +
1.5 &mu;l dNTPs (20 &mu;M ;Fermentas)
 +
0.5 &mu;l Pfu turbo polymerase (KNGiE)
 +
2.5 &mu;l Pfu Turbo Buffer (Fermentas)
 +
2.5 &mu;l MgSO<sub>4</sub> (20 &mu;M; Fermentas)
 +
1 &mu;l DNA template
 +
16.5 &mu;l MQ water</pre></li>
 +
<li>proprer mixture 2:
 +
<pre>0.25 &mu;l primer 1 (50 nM; Oligo.pl)
 +
0.25 &mu;l primer 2 (50 nM; Oligo.pl)
 +
1.5 &mu;l dNTPs (20 &mu;M ;Fermentas)
 +
0.5 &mu;l Pfu turbo polymerase (KNGiE)
 +
2.5 &mu;l Pfu Turbo Buffer (Fermentas)
 +
2.5 &mu;l MgSO<sub>4</sub> (20 &mu;M; Fermentas)
 +
2 &mu;l DNA template
 +
15.5 &mu;l MQ water</pre></li>
 +
<li>Negative control: the same as proper mixture 1, the only distinction is lack of the DNA template.</li>
 +
</ol></ul>
 +
<ul>
 +
<li>Program:</li>
 +
</ul>
 +
<br/>
 +
<p>p53 (detailed destription is <a href="https://2009.igem.org/Team:Warsaw/Calendar-Main/9_July_2009">here</a></p>
 +
<br/>
 +
<p>Results:</p>
 +
<ul>
 +
<li>Clean-up the PCR products</li>
 +
</ul>
 +
<p>Procedure:</p>
 +
<ul>
 +
<li>DNA was purified using the A&A clean-up kit. Detailed procedure is described <a href="http://www.aabiot.com/products/dna_purification/dna_fragments/clean_up/protocol_clean_up.pdf">here</a></li>
 +
<li>After clean-up 1 &mu;l of purified PCR product was loaded into gel and photographed.</li></ul>
 +
<br/>
 +
<center>
 +
<img src="https://static.igem.org/mediawiki/2009/2/26/PCR_reaction-p53_13_07_09.png" width="45%" height="45%">
 +
</center>
 +
<p><div style="text-align: center;">verification of PCR reaction and clean-up procedure</div><p>
 +
<p><b>Comment:</b></p>
 +
<p> Unfortunatelly reaction 1 (2 &mu;l of DNA matrix) was abortive - there is no product in the gel.</p>
 +
<br/>
 +
<p>Task 2:</p>
 +
<ul>
 +
<li>Restriction digest of p53 coding sequence.</li>
 +
</ul>
 +
<p>Methods:</p>
 +
<ul>
 +
<li>Control digest using PvuII</li>
 +
<ul><li>Reaction mixture composition:  
 +
<pre>1 &mu;l purified PCR product
 +
0.5 &mu;l PvuII (Fermentas)
 +
2 &mu;l Buffer Green (Fermentas)
 +
15.5 &mu;l MQ water</pre></li></ul>
 +
<li>Digest for subsequent cloning using XbaI</li>
 +
<ul><li>Reaction mixture composition:  
 +
<pre>10 &mu;l purified PCR product
 +
1 &mu;l XbaI (Fermentas)
 +
5 &mu;l Buffer Tango (Fermentas)
 +
34.5 &mu;l MQ water</pre></li></ul>
 +
<li>Both reaction were perform in the same condition:</li>
 +
</ul>
 +
<p>Program:</p>
 +
<p>digest:</p>
<pre>
<pre>
1. 37&deg;C - 3 hours
1. 37&deg;C - 3 hours
Line 47: Line 88:
3. 4&deg;C - hold
3. 4&deg;C - hold
</pre>
</pre>
-
 
+
<br/><center>
-
[[image:P53_control_digest_13_07_09.png‎|thumb|300px|center|control digest of p53 with PvuII ]]
+
<img src="https://static.igem.org/mediawiki/2009/6/63/P53_control_digest_13_07_09.png" heigth="50%" width="50%"><center>
-
 
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<div style="text-align: central;">control digest of p53 with PvuII</div>
-
Restriction pattern confirm that sequence is correct
+
<br/>
-
 
+
<p><b><Comment</b></p>
-
*Purification of digested products via gel-out
+
<p>Restriction pattern confirms that sequence is correct</p>
-
 
+
<ul>
-
Procedure:
+
<li>Purification of digested products via gel-out</li>
-
*Fragments of agarose gel were carefully cut out and in the next step DNA was extracted from the gel using the A&A gel-out kit. Detailed procedure is described [http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf here]
+
</ul>
-
 
+
<br/>
-
*Quantification of amount of p53 DNA after restriction digest:
+
<p>Procedure:<p/>
-
 
+
<ul>
-
1 &mu;l of the digest mixture was diluted to 10 &mu;l and loaded into the gel.
+
<li>Fragments of agarose gel were carefully cut out and in the next step DNA was extracted from the gel using the A&A gel-out kit. Detailed procedure is described <a href="http://aabiot.com/products/dna_purification/dna_fragments/gel_out/protocol_gel_out.pdf">here</a></li>
-
 
+
<li>Quantification of amount of p53 DNA after restriction digest:</li>
-
[[image:P53_PCR_after_digest_13_07_09.png‎ |thumb|center|200px|PCR product after digest loaded into the gel]]
+
</ul>
-
 
+
<p>1 &mu;l of the digest mixture was diluted to 10 &mu;l and loaded into the gel.</p>
-
Task 3:
+
<br/><center>
-
 
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<img src="https://static.igem.org/mediawiki/2009/4/47/P53_PCR_after_digest_13_07_09.png" width="50%" heigth="50%"><center>
-
*Cloning p53 coding sequence to pKS plasmid
+
<p><div style="text-align: central;">PCR product after digest loaded into the gel</div></p>
-
Methods:
+
<br/>
-
*Ligation mixture composition: 14 &mu;l digested p53, 1.5 &mu;l digested pKS, 5 &mu;l ligation buffer (Invitrogen), 1 &mu;l ligase T4
+
<p>Task 3:</p>
-
* Duration of ligation was about 12 hours   
+
<br/>
 +
<ul>
 +
<li>Cloning p53 coding sequence to pKS plasmid</li>
 +
</ul>
 +
<p>Methods:</p>
 +
<ul>
 +
<li>Ligation mixture composition:  
 +
<pre>14 &mu;l digested p53
 +
1.5 &mu;l digested pKS
 +
5 &mu;l ligation buffer (Invitrogen)
 +
1 &mu;l ligase T4</pre></li>
 +
<li>Duration of ligation was about 12 hours</li>
 +
</ul>
 +
</html>  
Line 76: Line 130:
'''Franek'''
'''Franek'''
-
 
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__NOTOC__
<br>
<br>
Tasks:
Tasks:
Line 98: Line 152:
* Positive selection will be made according to fluorescence under UV light
* Positive selection will be made according to fluorescence under UV light
<!-- TU PISZ CO CHCESZ! -->
<!-- TU PISZ CO CHCESZ! -->
 +
<html><h3><div style="text-align: center;">Making of the RBS-cI</div></h3>
 +
<h4>Jarek</h4>
 +
<br />
 +
<p>Tasks:</p>
 +
<ul>
 +
<li>Preparing 20 liquid cultures from colonies that grew after transformation and incubationg them in 37C degree.</li>
 +
<li>Isolation of plasmid DNA from liquid cultures.</li>
 +
<li>Digestion of isolated DNA with EcoRI and PstI endonucleases.</li>
 +
<html>
<html>
Line 110: Line 173:
<p>Methods (the bulk technique):</p>
<p>Methods (the bulk technique):</p>
<ul>
<ul>
-
<li>Plasmid digest mix was prepared as follows: 2&mu;l Tango buffer (Fermentas), 3&mu;l pKSII+ plasmid, 2&mu;l XbaI enzyme, 2&mu;l SmaI enzyme, the solution was topped up with H2O to the final volume of 20 ul.</li>
+
<li>Plasmid digest mix was prepared as follows:  
-
<li>mgtc promoter digest mix was prepared as follows: 2&mu;l Tango buffer (Fermentas), 10&mu;l purified gene, 2&mu;l XbaI enzyme, the solution was topped up with H2O to the final volume of 20 ul.</li>
+
<pre>2&mu;l Tango buffer (Fermentas)
 +
3&mu;l pKSII+ plasmid
 +
2&mu;l XbaI enzyme
 +
2&mu;l SmaI enzyme</pre>
 +
the solution was topped up with H2O to the final volume of 20 ul.</li>
 +
<li>mgtc promoter digest mix was prepared as follows
 +
<pre>2&mu;l Tango buffer (Fermentas)
 +
10&mu;l purified gene
 +
2&mu;l XbaI enzyme</pre>
 +
the solution was topped up with H2O to the final volume of 20 ul.</li>
<li>The digest was kept for 3h at 37&deg;C, and then the enzyme was inactivated for 15min. at 80&deg;C.</li>
<li>The digest was kept for 3h at 37&deg;C, and then the enzyme was inactivated for 15min. at 80&deg;C.</li>
-
<li><p>The ligation mix was prepared as follows: both inactivated digests were mixed together and topped with 5&mu;l of 30% PEG, 5&mu;l 10mM ATP, 1,2&mu;l Tango buffer (Fermentas) and 2&mu;l of T4 ligase (Fermentas). The ligation was carried out in 18&deg;C overnight (~18h) and then inactivated for 10min. at 65&deg;C.</li>
+
<li><p>The ligation mix was prepared as follows:
 +
<pre>both inactivated digests were mixed together and topped with 5&mu;l of 30% PEG
 +
5&mu;l 10mM ATP
 +
1,2&mu;l Tango buffer (Fermentas)
 +
2&mu;l of T4 ligase (Fermentas)</pre>
 +
The ligation was carried out in 18&deg;C overnight (~18h) and then inactivated for 10min. at 65&deg;C.</li>
</var>
</var>
<br />
<br />
-
 
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<h3>Cloning of hly gene into pKSII+ vector</h3>
 +
<h4>Kama</h4>
 +
<ul>
 +
<li>chemocompetent E. coli dH5&alpha; were incubated on ice for 15 minutes</li>
 +
<li>ligation mixture was added</li>
 +
<li>bacteria were incubated with DNA on ice for 30 minutes</li>
 +
<li>heat shock was conducted (1 minute 42&deg;C)</li>
 +
<li>bacteria were incubated on ice for 3 minutes</li>
 +
<li>After the heat shock 750&mu;l of SOB medium was added</li>
 +
<li>Mixture with bacteria was incubated for 1 hour in 37&deg;C</li>
 +
<li>100&mu;l of mixture was plated on medium containing ampicillin, X-Gal and IPTG (diluted 10* and without dilution)</li>
 +
</ul>
 +
</p>
 +
</ul>
 +
</li>
 +
</p>
 +
</ul>
 +
</li>
 +
</ul>
 +
</li>
 +
</ul>
 +
</li>
</html>
</html>
 +
<h3> <div style="text-align: center;">Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 K177012] operon1_part2 </div></h3>
 +
 +
<h4>Ania</h4>
 +
 +
Tasks:
 +
 +
* Set up a liquid culture from traqnsformants containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid. 
 +
 +
* Alkaline lysis of bacterial cultures to obtain plasmids containing following bricks:
 +
[http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid. 
 +
 +
* Digest of [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid (XbaI/PstI = prospective insert)
 +
* Digest of [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 BBa_B0032 - RBS.3] on the pSB1A2 ampicillin resistant plasmid (SpeI/PstI = prospective  vector)
 +
 +
Results:
 +
 +
* Fail. Digestion mix was incubated overnight at room temperature, this might be the reason for the unproper digestion.
 +
 +
===<div style="text-align: center;">Isolation of BioBricks from 2008 and 2009 Kit Plates</div>===
 +
 +
'''Monika'''
 +
 +
Task 1
 +
*Another attempt to isolate GFP coding device switched on by IPTG - [http://partsregistry.org/Part:BBa_I763004 <span style="color: blue;">BBa_I763004</span>]  from 2008 Kit Plate 1017 well G5
 +
 +
 +
Methods
 +
*2008 Kit: isolation of DNA from selected wells (two punched paper spots) with 8ul of TE (prodedure described
 +
[http://partsregistry.org/Help:IGEM_08_DNA_distribution <span style="color: blue;"> here</span>])
 +
*Transformation of chemocompetent cells (prepared by Franek and Ania) with 3ul of DNA solution
 +
*Planting on LB-agar medium supplemented with ampicillin and kanamycin
 +
 +
Results
 +
*Will be determined tomorrow
 +
Task 2
 +
*prepare culture of bacteria to isolate promoter lambda (cI regulated) with RFP reporter [http://partsregistry.org/Part:BBa_I763007 <span style="color: blue;">BBa_I763007</span>]

Latest revision as of 21:35, 19 September 2009


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Cloning of p53 coding sequence

Marcin


Task 1:

  • Prepare PCR reaction to amplified p53 coding sequence.

Methods:

  • DNA template dilution:

1 &mi;l of plasmid solution (isolated in 11.07.09) was taken and diluted with 99 mi&;l of MQ water

  • PCR mixture composition:
    1. proper mixture 1: 
      0.25 μl primer 1 (50 nM; Oligo.pl) 
0.25 μl primer 2 (50 nM; Oligo.pl)
1.5 μl dNTPs (20 μM ;Fermentas)
0.5 μl Pfu turbo polymerase (KNGiE)
2.5 μl Pfu Turbo Buffer (Fermentas)
2.5 μl MgSO4 (20 μM; Fermentas)
1 μl DNA template
16.5 μl MQ water
    2. proprer mixture 2: 
      0.25 μl primer 1 (50 nM; Oligo.pl)
0.25 μl primer 2 (50 nM; Oligo.pl)
1.5 μl dNTPs (20 μM ;Fermentas)
0.5 μl Pfu turbo polymerase (KNGiE)
2.5 μl Pfu Turbo Buffer (Fermentas)
2.5 μl MgSO4 (20 μM; Fermentas)
2 μl DNA template
15.5 μl MQ water
    3. Negative control: the same as proper mixture 1, the only distinction is lack of the DNA template.
  • Program:

p53 (detailed destription is here


Results:

  • Clean-up the PCR products

Procedure:

  • DNA was purified using the A&A clean-up kit. Detailed procedure is described here
  • After clean-up 1 μl of purified PCR product was loaded into gel and photographed.

verification of PCR reaction and clean-up procedure

Comment:

Unfortunatelly reaction 1 (2 μl of DNA matrix) was abortive - there is no product in the gel.


Task 2:

  • Restriction digest of p53 coding sequence.

Methods:

  • Control digest using PvuII
    • Reaction mixture composition: 
      1 μl purified PCR product
0.5 μl PvuII (Fermentas)
2 μl Buffer Green (Fermentas)
15.5 μl MQ water
  • Digest for subsequent cloning using XbaI
    • Reaction mixture composition: 
      10 μl purified PCR product
1 μl XbaI (Fermentas)
5 μl Buffer Tango (Fermentas)
34.5 μl MQ water
  • Both reaction were perform in the same condition:

Program:

digest:

1. 37°C - 3 hours
2. 80°C - 15 minutes
3. 4°C - hold

control digest of p53 with PvuII

Restriction pattern confirms that sequence is correct

  • Purification of digested products via gel-out

Procedure:

  • Fragments of agarose gel were carefully cut out and in the next step DNA was extracted from the gel using the A&A gel-out kit. Detailed procedure is described here
  • Quantification of amount of p53 DNA after restriction digest:

1 μl of the digest mixture was diluted to 10 μl and loaded into the gel.


PCR product after digest loaded into the gel


Task 3:


  • Cloning p53 coding sequence to pKS plasmid

Methods:

  • Ligation mixture composition: 
    14 μl digested p53
1.5 μl digested pKS
5 μl ligation buffer (Invitrogen)
1 μl ligase T4
  • Duration of ligation was about 12 hours


Creating devices to test promoters in E. coli strains (devices [http://partsregistry.org/Part:BBa_K177024 BBa_K177024] and [http://partsregistry.org/Part:BBa_K177025 BBa_K177025])

Franek


Tasks:

  • Digestion of [http://partsregistry.org/Part:BBa_R0010 BBa_R0010 - lacI regulated promoter], [http://partsregistry.org/Part:BBa_R0080 BBa_R0080 - AraC regulated promoter] and [http://partsregistry.org/Part:BBa_E0840 E0840 – RBS + GFP + Terminator]
  • Ligation of [http://partsregistry.org/Part:BBa_R0010 lacI regulated promoter] and [http://partsregistry.org/Part:BBa_R0080 AraC regulated promoter] with [http://partsregistry.org/Part:BBa_E0840 E0840]


Methods:

  • DNA containing [http://partsregistry.org/Part:BBa_R0080 pAraC] and [http://partsregistry.org/Part:BBa_R0010 placI] was digested with SpeI and PstI enzymes. Digestion mix contained 10µl of extracted DNA, 2µl of Fermentas Tango buffer, 0.5µl of each enzyme and water added to obtain 20µl total volume. Both mixes were incubated for 3h at 37°C.
  • DNA containing [http://partsregistry.org/Part:BBa_E0840 E0840] was digested with XbaI and PstI enzymes. Digestion mix contained 26µl of extracted DNA, 3µl of Fermentas Tango buffer, 0.5µl of each enzyme. Two identical mixes created this way were incubated for 3h at 37°C.
  • Enzymes in all 4 mixes were inactivated through incubation at 80°C for 20 min.
  • Two ligation mixes were prepared. Each contained 30µl of [http://partsregistry.org/Part:BBa_R0080 pAraC]/ [http://partsregistry.org/Part:BBa_R0010 placI] digestion mix, 20µl of [http://partsregistry.org/Part:BBa_E0840 E0840] mix, 5µl of dNTPs and 2µl of ligase. Both were left at 16°C for over night incubation.


Results:

  • Positive selection will be made according to fluorescence under UV light

Making of the RBS-cI

Jarek


Tasks:

  • Preparing 20 liquid cultures from colonies that grew after transformation and incubationg them in 37C degree.
  • Isolation of plasmid DNA from liquid cultures.
  • Digestion of isolated DNA with EcoRI and PstI endonucleases.

    • Cloning of the mgtc promoter into the pKSII+ plasmid

    Kamil


    Tasks:

    • Plasmid assembly

    Methods (the bulk technique):

    • Plasmid digest mix was prepared as follows: 
      2μl Tango buffer (Fermentas)
3μl pKSII+ plasmid
2μl XbaI enzyme
2μl SmaI enzyme
      the solution was topped up with H2O to the final volume of 20 ul.
    • mgtc promoter digest mix was prepared as follows 
      2μl Tango buffer (Fermentas)
10μl purified gene
2μl XbaI enzyme
      the solution was topped up with H2O to the final volume of 20 ul.
    • The digest was kept for 3h at 37°C, and then the enzyme was inactivated for 15min. at 80°C.

    • The ligation mix was prepared as follows: 

      both inactivated digests were mixed together and topped with 5μl of 30% PEG
5μl 10mM ATP
1,2μl Tango buffer (Fermentas)
2μl of T4 ligase (Fermentas)
      The ligation was carried out in 18°C overnight (~18h) and then inactivated for 10min. at 65°C.

      • Cloning of hly gene into pKSII+ vector

      Kama

      • chemocompetent E. coli dH5α were incubated on ice for 15 minutes
      • ligation mixture was added
      • bacteria were incubated with DNA on ice for 30 minutes
      • heat shock was conducted (1 minute 42°C)
      • bacteria were incubated on ice for 3 minutes
      • After the heat shock 750μl of SOB medium was added
      • Mixture with bacteria was incubated for 1 hour in 37°C
      • 100μl of mixture was plated on medium containing ampicillin, X-Gal and IPTG (diluted 10* and without dilution)

Construction of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K177012 K177012] operon1_part2

Ania

Tasks:

  • Set up a liquid culture from traqnsformants containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid.
  • Alkaline lysis of bacterial cultures to obtain plasmids containing following bricks:

[http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid.

  • Digest of [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012 - lacI repressor] on the pSB1A2 ampicillin resistant plasmid (XbaI/PstI = prospective insert)
  • Digest of [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 BBa_B0032 - RBS.3] on the pSB1A2 ampicillin resistant plasmid (SpeI/PstI = prospective vector)

Results:

  • Fail. Digestion mix was incubated overnight at room temperature, this might be the reason for the unproper digestion.

Isolation of BioBricks from 2008 and 2009 Kit Plates

Monika

Task 1

  • Another attempt to isolate GFP coding device switched on by IPTG - [http://partsregistry.org/Part:BBa_I763004 BBa_I763004] from 2008 Kit Plate 1017 well G5


Methods

  • 2008 Kit: isolation of DNA from selected wells (two punched paper spots) with 8ul of TE (prodedure described

[http://partsregistry.org/Help:IGEM_08_DNA_distribution here])

  • Transformation of chemocompetent cells (prepared by Franek and Ania) with 3ul of DNA solution
  • Planting on LB-agar medium supplemented with ampicillin and kanamycin


Results

  • Will be determined tomorrow

Task 2

  • prepare culture of bacteria to isolate promoter lambda (cI regulated) with RFP reporter [http://partsregistry.org/Part:BBa_I763007 BBa_I763007]



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