IGEM:Paris Bettencourt 2012/Notebooks/Semantic group/day by day//2012/07/17: Difference between revisions

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As positive control, I transform in the same condition the R0011 which has already been shown to be a good plasmid (pSB1A2)
As positive control, I transform in the same condition the R0011 which has already been shown to be a good plasmid (pSB1A2)
As negative control, I just spread the NEB turbo without adding any DNA.
As negative control, I just spread the NEB turbo without adding any DNA.
'''EDIT : there is nothing for K228006, because I plate on the wrong ATB plate.'''


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Revision as of 06:38, 19 July 2012

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BioBrick R0011 (pLac)

  • Miniprep (Fermentas) of the 10mL of O/N Culture.
  • Plasmid recovered in 50µL of Water
  • NanoDrop :
    • 73 ng/µL
    • DO260/280 = 1,96
  • Store at -20°C

Biobricks retrieval

Parts number Plasmid Name Distribution/Plate/Well
BBa_K228001 supD (tRNA amber supressor gene) pSB1A2 2012/4/8K
BBa_B1006 terminator pSB1AK3 2012/1/4H
  • I transform them in NEB turbo, competent cells, along with the miniprep of BB R0011 (pSC001)

I transform 0.5 µL of plasmid in 20 µL of cells.

    • Let 30' on ice
    • heat shock 42°C
    • 2' on ice
    • add 200 µL of warm LB (pre-heated at 42°C)
    • shaking at 37°C for 1h
    • Spread 20 µL and 180 µL in LB+Amp or L+Amp+Kan (K228001) plates
    • O/N 37°C

As positive control, I transform in the same condition the R0011 which has already been shown to be a good plasmid (pSB1A2) As negative control, I just spread the NEB turbo without adding any DNA.

EDIT : there is nothing for K228006, because I plate on the wrong ATB plate.

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