IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/09/07: Difference between revisions
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== | ==Colicin Assay== | ||
[[Image:SGdNEB08C2060912.jpg |300px]] | [[Image:SGdNEB08C2060912.jpg |300px]] |
Revision as of 04:32, 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