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Materials
You will need:
Extra Notes
Do not keep cells out for too long. It is vital that the cells undergo a rapid temperature change (from very cold to very hot suddenly) in order for crack pores in the cell wall.
Procedure
- Thaw DH5α cells on ice for 10 minutes.
- Transfer 50μL of DH5α cells to chill, sterile polypropylene tubes.
- Transfer 1μL of DNA into the tubes with the cells.
- Incubate on ice for 30 minutes.
- Gently transfer tubes to 42°C water bath for 90s (for heat shock)
- Transfer tubes back on ice for 2 minutes.
- Add 800μL of luria broth into each tube.
- Incubate at 37°C for 1 hour in a shaker.
- Plate 200μL of each sample on nutrient agar plates. (Be sure to choose the plates with the antibiotic that your E. Coli is resistant to.)
- Spread the sample evenly by plating with sterilized glass beads. Empty the plate of glass beads afterwards.
- Incubate overnight at 37°C.
Purpose
To insert plasmids into E. Coli.
References
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We would like to take a moment to thank all of our sponsors for their very generous donations, as we could not have done this without your help!
We would also like to thank and acknowledge:
Our Advisors
Marc Facciotti
Ilias Tagkopoulos
Technical Guidance
David Larsen
Andrew Yao
Visiting iGEMer
Jia Li of Zhejiang University (TEAM ZJU-China)
cI Promoter Screen
Drew Endy - Stanford
Thomas Schneider - NIH
Want to sponsor us? Send an email to mtfacciotti@ucdavis.edu to discuss various ways you can help! :)
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