Team:Warsaw/Calendar-Main/19 May 2008

From 2008.igem.org

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<html><h3>PCRs for fusions</h3>
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<html><h3>PCRs for fusions<br>
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<h4>Michał K.:</h4>
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Michał K.</h3>
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<li>Electrophoresis to estimate the concentration of isolated DNA.</li></ol></p>
<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/>
<img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/>
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<h3>1-DNA ladder; 2-PCR product-translation fusion</h3>
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<var>1-DNA ladder; 2-PCR product-translation fusion</var>
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<h4>Michał L., Ewa, Marcin:</h4>
<h4>Michał L., Ewa, Marcin:</h4>
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<ol><li>Overnight inoculation and induction:</li>  
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<p>Overnight inoculation and induction:<br>
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<p><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> + 0,5 mM IPTG</p>
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<a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">Top10</a> + 0,5 mM IPTG<br>
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<p>pMPM-AID + Glc + 0,01% L-Ara</p>
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pMPM-AID + Glc + 0,01% L-Ara<br>
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<p><a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + Glc + 0,5 mM IPTG</p>
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<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + Glc + 0,5 mM IPTG<br>
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Revision as of 14:12, 9 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).
  3. Electrophoresis to estimate the concentration of isolated DNA.

1-DNA ladder; 2-PCR product-translation fusion

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