Team:Warsaw/Calendar-Main/19 May 2008

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Latest revision as of 16:37, 28 October 2008


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Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.

PCR - translational fusion: AID + T7 RNA-polymerase
Primers:
- AIDlNrH
- T7pXbSal
Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translational fusion
Annealing temperature : 73°C
Elongation time: 4 minutes
20 cycles
Gel electrophoresis and isolation of proper band (3300 bp). Fig. 1.
Electrophoresis to estimate the concentration of isolated DNA.

Fig. 1. 1-DNA ladder; 2-PCR product-translational fusion

Rifampicin test #1

Michał L., Ewa, Marcin

Overnight culture of the following strains:
Top10
Top10 + 0.5 mM IPTG
and Top10 carrying:
pMPM-T5-AID + Glc
pTrc99a-AID + Glc
pMPM-T5-AID + 0.01% L-Ara
pTrc99a-AID + 0.5 mM IPTG

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-<html>
+<html><h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPMT5-AID+T7</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a></h3>
+<h4>Michał K.</h4>
+<ol>
+<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>translational fusion</a>: AID + T7 RNA-polymerase<br>
-<p>Michał K.:</p>
+Primers: <br>
-<p>1. Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/16_May_2008">16 May</a> PCR's) and DNA <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> from proper bands (600 bp for AID lane and 2700 bp for T7 lanes).</html>
+<ul>
+<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDlNrH">AIDlNrH</a></li>
+<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#T7pXbSal">T7pXbSal</a></li>
+</ul>
+Template DNA: purified PCR products from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/16_May_2008">16 May</a> - AID and T7 RNA-polymerase for <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>translational fusion</a> <br>
+Annealing temperature : 73&deg;C <br>
+Elongation time: 4 minutes <br>
+cycles <br></li>
+<li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of proper band (3300 bp). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/19_May_2008#fig1">Fig. 1</a>.</li>
+<li>Electrophoresis to estimate the concentration of isolated DNA.</li></ol></p>
+<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=190/></a>
+<var><b>Fig. 1.</b> 1-DNA ladder; 2-PCR product-translational fusion</var>
+<h3>Rifampicin test #1</h3>
+<h4>Michał L., Ewa, Marcin</h4>
+<p>Overnight culture of the following strains:<br>
+<a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a><br>
+<a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> + 0.5 mM IPTG<br>
+and <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> carrying:<br>
+<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5-AID</a> + Glc <br>
+<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + Glc <br>
+<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5-AID</a> + 0.01% L-Ara<br>
+<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + 0.5 mM IPTG<br>
+</html>
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