Team:Warsaw/Calendar-Main/19 May 2008

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<html><h4>Michał K.:</h4>
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<html><h3>PCRs for fusions</h3>
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<h4>Michał K.:</h4>
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Revision as of 11:20, 5 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).

Michał L., Ewa, Marcin:

  1. Overnight inoculation and induction:

    2. Top10 + 0,5 mM IPTG

    pMPM-AID + Glc + 0,01% L-Ara

    pTrc99a-AID + Glc + 0,5 mM IPTG

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