Team:Warsaw/Calendar-Main/26 June 2008

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Latest revision as of 14:02, 28 October 2008


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Change of the reporter from pZC320 with B-galactosidase to GFP or RFP

Piotr, Weronika

Isolation of pZC30.
Inoculation of E.coli TOP10 carrying pSB1A2 standard plasmids carrying parts: BBa_E0840(GFP genetrator) and BBa_J04450 (RFP generator).

Preparation of constructs: OmpA_alpha and OmpA_omega

Michał K.

Repetition of PCRs, gel-out isolations and digests from 17 June.

Preparation of alpha-A and omega-A fusions

Michał L., Ewa, Marcin

Polymerase Chain Ligation (PCL)

Product	Templates	Primers	Product length
Alpha-A fusion	Alpha-linker + linker-A	AlphaL+SacI + AP+NotI	1100 bp
Omega-A fusion	Omega-linker + linker-A	OmegaL+SacI + AP+NotI	750 bp

PCL program for fusion A with alpha and omega

Temperature	Time
94°C	4:00
94°C	0:30	28 cycles
48°C-62°C	0:45
68°C	1:30
68°C	10:00
4°C	infinite

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+<html>
+<html>
-<p>'''Preparation of pMPMT5-AID+AIDT7 construct''' <br>
+<h3>Change of the reporter from <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a> with B-galactosidase to GFP or RFP</h3>
-. Transformation of E. coli TOP10  strain with ligation from previous day <br>
+<h4>Piotr, Weronika</h4>
-. Plating transformants on LB + tetracycline</p>
+<ol>
+<li>Isolation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC30</a>.</li>
+<li>Inoculation of E.coli TOP10 carrying <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A2>pSB1A2</a> standard plasmids carrying parts: <A href=http://partsregistry.org/wiki/index.php?title=Part:BBa_E0840>BBa_E0840</a>(GFP genetrator) and <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450>BBa_J04450</a> (RFP generator).
+</li></ol>
+<h3>Preparation of constructs: OmpA_alpha and OmpA_omega</h3><h4>Michał K.</h4>
+<p>Repetition of PCRs, gel-out isolations and digests from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_June_2008">17 June.</a></p>
+<h3>Preparation of alpha-A and omega-A fusions</h3><h4>Michał L., Ewa, Marcin</h4>
+<b><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcl">Polymerase Chain Ligation (PCL)</a></b>
+<table id="result">
+<tr><th>Product</th><th>Templates</th><th>Primers</th><th>Product length</th></tr>
+<tr><th>Alpha-A fusion</th><td>Alpha-linker + linker-A</td><td>AlphaL+SacI + AP+NotI </td><td>1100 bp</td></tr>
+<tr><th>Omega-A fusion</th><td>Omega-linker + linker-A</td><td>OmegaL+SacI + AP+NotI </td><td>750 bp</td></tr>
+</table>
+<br><br>
+<b>PCL program for fusion A with alpha and omega</b><br>
+<table id="result">
+<tr><th>Temperature</th><th>Time</th></tr>
+<tr><td>94&deg;C</td><td>4:00</td></tr>
+<tr><td>94&deg;C</td><td>0:30</td><td rowspan="3">28 cycles</td></tr>
+<tr><td>48&deg;C-62&deg;C</td><td>0:45</td></tr>
+<tr><td>68&deg;C</td><td>1:30</td></tr>
+<tr><td>68&deg;C</td><td>10:00</td></tr>
+<tr><td>4&deg;C</td><td>infinite</td></tr>
+</table>
+</html>
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