Smolke:Protocols/Transformation

From OpenWetWare

< Smolke:Protocols
Revision as of 14:14, 15 October 2009 by Josh K. Michener (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search

Home        Contact        Internal        Protocols        Lab Members        Publications        Research       


  • For each sample
    • 1 tube of electrocompetent cells (keep covered in ice at all time)
    • 1 electroporation cuvette (keep on ice)
    • 1 1.5-ml microfuge tube
    • 1 mL SOC
    • 1 LB plate with appropriate antibiotics
  • Gene Pulser setup
    • Exponential
    • 2000 V voltage
    • 25 uF capacitance
    • 200 ohms resistance
    • 1 mm cuvette
  • If transforming non-purified, non-concentrated ligation product, use 0.5-1.0 ul per sample.
  • Time constant should be at least 4.5, generally between 4.7 and 5.0.
  • If sample sparks or has a low time constant, lowering the volume of ligation product is generally more useful than increasing the volume.
Personal tools