Team:Warsaw/Calendar-Main/19 May 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

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