Team:Warsaw/Calendar-Main/19 May 2008

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(Difference between revisions)
Line 32: Line 32:
<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>
+
<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/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>
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/></a>
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/></a>

Revision as of 15:06, 12 October 2008

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PCRs for fusions

Michał K.

  1. PCR - translational fusion: AID + T7 RNA-polymerase
    Primers:
    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
  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-T5-AID + Glc + 0.01% L-Ara
pTrc99a-AID + Glc + 0.5 mM IPTG
Fig. 1. 1-DNA ladder; 2-PCR product-translational fusion


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