AlanFung:Protocol/Plasmid DNA amplification

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Revision as of 20:49, 20 September 2010 by >Alan6017518 (Created page with '==Plasmid DNA Amplification== ==For pRFP-C-RS based HuSH vector from Origene== ==Dilution of Plasmid solution== *Add 50ul of dH2O into each tube *Vortex the tubes to resuspend t…')
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Plasmid DNA Amplification

For pRFP-C-RS based HuSH vector from Origene

Dilution of Plasmid solution

  • Add 50ul of dH2O into each tube
  • Vortex the tubes to resuspend the DNA
  • Pipette 1ul of this soultion to another tube
  • Add 99ul H2O Concentration of the DNA solution should be 1ng/ul
  • Notes: Store plasmid solution at -20C

Grow cells

  • Thaw transformation competent E.Coli cells on ice
  • Perform transformation with 1-2ul of diluted shRNA plasmid
  • Plate out the transformants on LB-Chloramphenicol plates
  • Incubate overnight at 37C, until colonies appear