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	9	2.66
Top10 pMPM-AID	none	15	2.5
Top10 pMPM-AID	0.1% Arabinose	42	2.27
Top10 pTRC-AID	none	12	2.41
Top10 pTRC-AID	0.5 mM IPTG	35	1.98

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

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

@@ Line 10: / Line 10: @@
 <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>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>
 <tr><th>Top10 pTRC-AID</th><td>none</td><td>12</td><td>2.41</td></tr>
-<tr><th>Top10 pTRC-AID</th><td>0.5 mM IPTG</td><td>35</td><<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>
-<br/>
+<br>
 <p>Conclusion: AID seems to work, induction of AID may reduces the growth rate</p>
-<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
+<h3>Preparation of <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID%2BAID-T7>pMPMT5-AID+AIDT7</a> construct</h3>
 <h4>Michał K.</h4>
 <p><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> for AID transcriptional fusion with translational fusion AID+T7 RNA-polymerase. <br>