Team:Warsaw/Calendar-Main/19 May 2008

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(Difference between revisions)
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<p>Overnight inoculation and induction:<br>
<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/igem_notebook.htm#top10">Top10</a> + 0.5 mM IPTG<br>
-
pMPM-AID + Glc + 0,01% L-Ara<br>
+
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>
+
<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + Glc + 0.5 mM IPTG<br>
<img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/>
<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>
<var>Fig. 1. 1-DNA ladder; 2-PCR product-translation fusion</var></html>

Revision as of 13:34, 11 October 2008

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PCRs for fusions

Michał K.

  1. PCR - translation fusion: AID + T7 RNA-polymerase
    Primers:
    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
  2. Gel electrophoresis and isolation of proper band (3300 bp). Fig. 1.
  3. 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


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