IGEM:Peking/2007/Count:Transformation

From OpenWetWare

< IGEM:Peking | 2007
Revision as of 23:11, 20 August 2007 by Anting Xu (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search
  1. Hold competent cells (from -80℃ refrigerator) on ice.
  2. Gently mix ligation product (10-15 μL) with cells, then bath in ice for 30 min.
  3. (Precisely!) Bath in 42℃ water for 90 sec.
  4. Hold again in ice for 2 min.
  5. Bath in 37℃ for 45 min. For plasmid pKO3 (and other temperature-sensitive strains), bath in 30℃ for 1 hour.
  6. Separate cells on petri-dishes, and cultivate them in 37℃ for 12 hours (for pKO3, 24 hours are necessary).
Personal tools