Team:Warsaw/Calendar-Main/19 May 2008

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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>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/></a>
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<var><b>Fig. 1.</b> 1-DNA ladder; 2-PCR product-translational fusion</var>
<h3>Rifampicin test</h3>
<h3>Rifampicin test</h3>
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<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/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 href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + 0.5 mM IPTG<br>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/></a>
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</html>
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<var><b>Fig. 1.</b> 1-DNA ladder; 2-PCR product-translational fusion</var></html>
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Revision as of 14:00, 26 October 2008

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Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

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.

Fig. 1. 1-DNA ladder; 2-PCR product-translational fusion

Rifampicin test

Michał L., Ewa, Marcin

Overnight culture of the following strains:
Top10
Top10 + 0.5 mM IPTG
and Top10 carrying:
pMPM-T5-AID + Glc
pTrc99a-AID + Glc
pMPM-T5-AID + 0.01% L-Ara
pTrc99a-AID + 0.5 mM IPTG

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