Team:Warsaw/Calendar-Main/19 May 2008

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Revision as of 11:37, 19 October 2008


	Attribution analyzer / Interactive list of topics

PCRs for fusions

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.

Rifampicin Test

Michał L., Ewa, Marcin

Overnight inoculation and induction:
Top10 + 0.5 mM IPTG
pMPM-T5-AID + Glc
pTrc99a-AID + Glc
pMPM-T5-AID + 0.01% L-Ara
pTrc99a-AID + 0.5 mM IPTG
Fig. 1. 1-DNA ladder; 2-PCR product-translational fusion

@@ Line 32: / Line 32: @@
 <p>Overnight inoculation and induction:<br>
 <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> + 0.5 mM IPTG<br>
-<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5-AID</a> + Glc + 0.01% L-Ara<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 + 0.5 mM IPTG<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>
 <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/></a>
 <var><b>Fig. 1.</b> 1-DNA ladder; 2-PCR product-translational fusion</var></html>