AU Biomaterials Design Lab:Protocols/Transformation Protocol: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(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...) |
|||
| Line 5: | Line 5: | ||
==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 10:51, 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
