Team:Warsaw/Calendar-Main/2 June 2008

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Rifampicin test #3

Piotr

StrainInducerNumber of colonies on rifampicinOD
TOP10none92.66
TOP10 pMPM-T5-AIDnone152.5
TOP10 pMPM-T5-AID0.1% Arabinose422.27
TOP10 pTrc99a-AIDnone122.41
TOP10 pTrc99a-AID0.5 mM IPTG351.98

Conclusion: AID seems to work, induction of AID may reduces the growth rate

Preparation of pMPMT5-AID+AIDT7 construct

Michał K.

PCR for AID transcriptional fusion with translational fusion AID+T7 RNA-polymerase.
Primers: AIDlNrH and T7pXbSal
Template DNA: translational fusion AID+T7 RNA-polymerase (pMPMT5+AID-T7)
Elongation temperature: 55°C
Time: 4 minutes
Number of cycles: 20 and 25

Blue/white and rifampicin test #1

Michał L., Ewa, Marcin

  1. Transformation of E. coli TOP10 with pZC320.
  2. Plating transformants on LB + ampicillin (30 μg/mL) + X-gal + IPTG.

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