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

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(Result)
Current revision (18:35, 24 September 2012) (view source)
(Colicin experiment)
 
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With the induction of IPTG we can still clearly see the clearing ring, which mean that the immunity protein did not work as expected.
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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

Image:090812.png

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
 
 
 
 
 

With the induction of IPTG we can still clearly see the clearing ring, which mean that the immunity protein did not work as expected.

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