IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/09/07

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==Colicin Assay==
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[[Image:SGdNEB08C2060912.jpg |300px]]
[[Image:SGdNEB08C2060912.jpg |300px]]

Revision as of 07:32, 17 September 2012

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Colicin Assay

Biobricks

We put pSG.008 (pLac+ColE2 immunity) and pSG.014 (ColE2 immunity) inside Biobricks vector (pSB1C3). All of them were cut with XbaI and PstI.

Protocol

PSB1C3 (-) BB+pSG.008 BB+pSG.014
H2O 6.5 uL - -
Insert 2 uL 2 uL 2 uL
Vector - 6.5 uL 6.5 uL
Ligase buffer 10x 1.0 uL 1.0 uL 1.0 uL
T4 ligase enzyme 0.5 uL 0.5 uL 0.5 uL
Total 10 uL 10 uL 10 uL

Transformation

We transformed 5 uL of each ligation product with heat shock transformation; +pSG.001 as a positive control

Gel

We ran the rest of the ligation product + 3 uL of green buffer; in a gel

Image:070912.png

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