Team:Hawaii/Protocols/Transformation E coli

From 2008.igem.org

Protocol

  1. Thawed cells on ice.
  2. Transformed 50 μl cells w/ 1 μl DNA
  3. Incubated on ice 30 min.
  4. Heat shocked 60 sec. in 42C water bath
  5. Added 250 μl SOC
  6. Incubated @ 37C for 1 hour with shaking (150 rpm)
  7. Plated 20-250 μl onto LB+antibiotic plates
  • Plate more if needed.

Notes

  • Adjust DNA amount as necessary. Transformation is saturated at >10ng DNA. DNA should not exceed 10% of cell volume (Re: SC).
  • Incubating 10 min. on ice after adding DNA and returning cells to ice for 2 min. after heat shock seems to improve transformation efficiency. -GK
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