IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/08/09

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Digestion pRG.037+pSG.014 (pSG.021)

We digested the minipreped pRG.037 (pLac inside pSB1A2, Denis)+pSG.014 (ColE2 immunity) with XbaI and PstI

Protocol

Colony 1 Colony 2 Colony 3 Colony 4 Colony 5
Plasmid (+/- 1 ug) 4 uL 6 uL 7 uL 4 uL 8 uL
H2O 28 uL 26 uL 25 uL 28 uL 24 uL
FD 10x Green Buffer 4 uL 4 uL 4 uL 4 uL 4 uL
XbaI 2 uL 2 uL 2 uL 2 uL 2 uL
PstI 2 uL 2 uL 2 uL 2 uL 2 uL
  • Incubate in 37°C, 400rpm for 1h15

Result

Colicin experiment

  • From the transformation result of pRG.037 (pLac inside pSB1A2, Denis) and pSG.014 (ColE2 immunity), we picked 5 colonies and grew them in liquid culture
  • The saturated cultures were diluted 100x in LB, then plated (20uL)
  • Drops of saturated ColE2 cells (5uL) were added to each plate
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