Team:Warsaw/Calendar-Main/2 June 2008

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<table id="result">
<table id="result">
<tr><th>Strain</th><th>Inducer</th><th>Number of colonies on rifampicin</th><th>OD</th></tr>
<tr><th>Strain</th><th>Inducer</th><th>Number of colonies on rifampicin</th><th>OD</th></tr>
-
<tr><th>Top10</th><td>none</td><td>100</td><td>2.66</td></tr>
+
<tr><th>Top10</th><td>none</td><td>9</td><td>2.66</td></tr>
<tr><th>Top10 pMPM-AID</th><td>none</td><td>15</td><td>2.5</td></tr>
<tr><th>Top10 pMPM-AID</th><td>none</td><td>15</td><td>2.5</td></tr>
<tr><th>Top10 pMPM-AID</th><td>0.1% Arabinose</td><td>42</td>><td>2.27</td></tr>
<tr><th>Top10 pMPM-AID</th><td>0.1% Arabinose</td><td>42</td>><td>2.27</td></tr>

Revision as of 12:33, 19 October 2008

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Results of rifampicin test and OD measurement

Piotr

><
StrainInducerNumber of colonies on rifampicinOD
Top10none92.66
Top10 pMPM-AIDnone152.5
Top10 pMPM-AID0.1% Arabinose422.27
Top10 pTRC-AIDnone122.41
Top10 pTRC-AID0.5 mM IPTG351.98

Conclusion: AID seems to work, induction of AID may reduces the growth rate

Preparation of pMPMT5-AID+AIDT7 construct

Michał K.

PCR for AID transcriptional fusion with translational fusion AID+T7 RNA-polymerase.
Primers: AIDlNrH and T7pXbSal
Template DNA: translational fusion AID+T7 RNA-polymerase (pMPMT5+AID-T7)
Elongation temperature: 55°C
Time: 4 minutes
Number of cycles: 20 and 25

Blue/white and rifampicin test

Michał L., Ewa, Marcin

  1. Transformation of E. coli TOP10 with pZC320.
  2. Plating transformants on LB + ampicillin (30 μg/mL) + X-gal + IPTG.

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