AU Biomaterials Design Lab:Protocols/Transformation Protocol

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(New page: ==Description== Transformation of PCR product into E. coli cells [http://tools.invitrogen.com/content/sfs/manuals/oneshotbl21_man.pdf Transformation Manual] ==materials== Materials Suppli...)
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Revision as of 12:37, 15 October 2012

Description

Transformation of PCR product into E. coli cells Transformation Manual

materials

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)

Protocol

  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
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