Team:Warsaw/Calendar-Main/19 May 2008

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Revision as of 13:24, 11 October 2008


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PCRs for fusions

Michał K.

PCR - translation fusion: AID + T7 RNA-polymerase
Primers:
- AIDlNrH
- T7pXbSal
Template DNA: purified PCR products from 16 May - AID and T7 RNA-polymerase for translation 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-AID + Glc + 0,01% L-Ara
pTrc99a-AID + Glc + 0,5 mM IPTG
Fig. 1. 1-DNA ladder; 2-PCR product-translation fusion

@@ Line 24: / Line 24: @@
 <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). Fig. 1.</li>
 <li>Electrophoresis to estimate the concentration of isolated DNA.</li></ol></p>
-<img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/>
-<var>Fig. 1. 1-DNA ladder; 2-PCR product-translation fusion</var>
@@ Line 35: / Line 34: @@
 pMPM-AID + Glc + 0,01% L-Ara<br>
 <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + Glc + 0,5 mM IPTG<br>
-</html>
+<img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/>
+<var>Fig. 1. 1-DNA ladder; 2-PCR product-translation fusion</var></html>
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