Team:Warsaw/Calendar-Main/19 May 2008

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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).</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>
<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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<var>1-DNA ladder; 2-PCR product-translation fusion</var>
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<var>Fig. 1. 1-DNA ladder; 2-PCR product-translation fusion</var>

Revision as of 09:20, 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.

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

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

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