AU Biomaterials Design Lab:Protocols/Transformation Protocol: Difference between revisions
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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 10:37, 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
- to PCR product, add 1 μL DPN1
- place reaction at 37 °C for 1 hour
store in freezer till ready for trasnformation
- prepare LB plates
- Equilibrate water bath to 42 °C and place the LB plates at 37 °C
- thaw onevial of cells on ice
- add 10 ng of DNA in 5 μL to the cells ( DO NOT MIX BY PIPETTING)
- heat shock the cells by incubating in 42 °C water bath for exactly 30 s
- place immediately on ice