AU Biomaterials Design Lab:Protocols/Transformation Protocol

From OpenWetWare

< AU Biomaterials Design Lab:Protocols
Revision as of 13:37, 15 October 2012 by Abigail E. Miller (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search


Transformation of PCR product into E. coli cells Transformation Manual


Materials Supplied by the User • Plasmid DNA (ready for transformation) • 42°C water bath • 37°C shaking and non-shaking incubator • Ice bucket with ice • Spectrophotometer to measure optical density of the cell cultures • Microcentrifuge tube rack (optional)


  1. to PCR product, add 1 μL DPN1
  2. place reaction at 37 °C for 1 hour

store in freezer till ready for trasnformation

  1. prepare LB plates
  2. Equilibrate water bath to 42 °C and place the LB plates at 37 °C
  1. thaw onevial of cells on ice
  2. add 10 ng of DNA in 5 μL to the cells ( DO NOT MIX BY PIPETTING)
  3. heat shock the cells by incubating in 42 °C water bath for exactly 30 s
  4. place immediately on ice
Personal tools