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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==materials== | ==materials== | ||
Materials Supplied by the User | 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 | |
| - | cell cultures | + | *Microcentrifuge tube rack (optional) |
| - | + | ||
==Protocol== | ==Protocol== | ||
Revision as of 13:51, 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
