IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/07/24: Difference between revisions

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==Entry title==
==Ligation of pSG.005 (pLac) and pSG.014 (ColE2 Immunity)==
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* As the control we ran also the digestion products and the ligation without insert
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[[Image:Sggel240712.jpg]]
[[Image:Sggel240712.jpg]]
*The gel looks strange but we still plated the ligation product in Cm plate.
==Transformation of BBa_K131000==
We transformed Colicin E2 full gene with SOS promoter to have the original ColE2 plasmid. The DNA was in the 2012 Kit Plate 3, well 22A. We plated in Kan plate and added Amp 20 uL because it has Amp and Kan resistance.
==pSG.006 Liquid Culture==
The day before we plated pSG.006 (pTet inside pSB4K5) on a plate with gradient antibiotic. We picked 5 big colonies closest to the center and grew each of them in 6ml LB+Kan.


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Revision as of 03:50, 25 July 2012

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Ligation of pSG.005 (pLac) and pSG.014 (ColE2 Immunity)

  • As the control we ran also the digestion products and the ligation without insert
PSG.005 (negative control) PSG.008 (ligation of pSG.005 and pSG.014)
Vector (30ng) 1 uL 1 uL
Insert (10x n vector) - 1.2 uL
Buffer 1uL 1 uL
Ligase T4 0.5 uL 0.5 uL
H2O 7.5 uL 6.3 uL

  • The gel looks strange but we still plated the ligation product in Cm plate.

Transformation of BBa_K131000

We transformed Colicin E2 full gene with SOS promoter to have the original ColE2 plasmid. The DNA was in the 2012 Kit Plate 3, well 22A. We plated in Kan plate and added Amp 20 uL because it has Amp and Kan resistance.

pSG.006 Liquid Culture

The day before we plated pSG.006 (pTet inside pSB4K5) on a plate with gradient antibiotic. We picked 5 big colonies closest to the center and grew each of them in 6ml LB+Kan.

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