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

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Line 27: Line 27:
  |[http://partsregistry.org/Part:BBa_K228001 BBa_K228001]
  |[http://partsregistry.org/Part:BBa_K228001 BBa_K228001]
  |supD (tRNA amber supressor gene)
  |supD (tRNA amber supressor gene)
  |pSB1AK3
  |pSB1A2
  |2012/4/8K
  |2012/4/8K
  |-  
  |-  
  |[http://partsregistry.org/Part:BBa_B1006 BBa_B1006]
  |[http://partsregistry.org/Part:BBa_B1006 BBa_B1006]
  |terminator
  |terminator
  |pSB1A2
  |pSB1AK3
  |2012/1/4H
  |2012/1/4H
  |-
  |-

Revision as of 10:50, 17 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.

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