Huang: DH5alpha transformation

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Contents

Overview

This protocol allows competent cell transformation.

Materials

  • DH5alpha competent cells
  • LB
  • LB + antibiotic plates
  • Plasmid DNA or DNA to transform the cells with

Procedure

  • 1. Set tubes of frozen cells on ice and let thaw very slowly
  • 2. Add 100ul thawed cells to new eppendorf
  • 3. Add 1-2ul DNA to cells
  • 4. Incubate on ice for 20 min
  • 5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C)
  • 6. Incubate on ice 2 min
  • 7. Add 900ul growth medium (LB) and shake 1 hour at 37C
  • 8. Plate about 200ul and grow overnight


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