Team:Warsaw/Calendar-Main/2 June 2008

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Results of rifampicin test and OD measurement

Piotr

><

Strain	Inducer	Number of colonies on rifampicin	OD
Top10	none	100	2.66
Top10 pMPM-AID	none	15	2.5
Top10 pMPM-AID	0.1% Arabinose	42	2.17
Top10 pTRC-AID	none	12
2.61
Top10 pTRC-AID	0.5 mM IPTG	35	1.98

Conclusion: AID seems to work, induction of AID 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

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

@@ Line 8: / Line 8: @@
 <table id="result">
 <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></tr><td>2.66</td></tr>
+<tr><th>Top10</th><td>none</td><td>100</td><td>2.66</td></tr>
-<tr><th>Top10 pMPM-AID</th><td>none</td><td>15</td></tr><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></tr><td>2.17</td></tr>
+<tr><th>Top10 pMPM-AID</th><td>0.1% Arabinose</td><td>42</td>><td>2.17</td></tr>
 <tr><th>Top10 pTRC-AID</th><td>none</td><td>12</td></tr><td>2.61</td></tr>
-<tr><th>Top10 pTRC-AID</th><td>0.5 mM IPTG</td><td>35</td></tr><td>1.98</td></tr>
+<tr><th>Top10 pTRC-AID</th><td>0.5 mM IPTG</td><td>35</td><<td>1.98</td></tr>
 </table>
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