Team:Warsaw/Calendar-Main/19 May 2008

From 2008.igem.org

(Difference between revisions)
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20 cycles <br></li>
20 cycles <br></li>
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<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>
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<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). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/19_May_2008#fig1">Fig. 1</a>.</li>
<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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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>
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<img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/></a>
<var><b>Fig. 1.</b> 1-DNA ladder; 2-PCR product-translational fusion</var></html>
<var><b>Fig. 1.</b> 1-DNA ladder; 2-PCR product-translational fusion</var></html>

Revision as of 23:10, 11 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-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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