IGEM:PennState/2008/PreparingElectrocompetentCells: Difference between revisions
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(New page: ='''Electrocompetent Cell Prep'''= ===<font color="red">Starter Culture</font>=== *Inoculate desired cell strain (DH5a) into 3 mL of LB and shake at 37C ===<font color="green">Pre...) |
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Latest revision as of 11:37, 22 May 2008
Electrocompetent Cell Prep
Starter Culture
- Inoculate desired cell strain (DH5a) into 3 mL of LB and shake at 37C
Prep
- OD the starter culture, use to calculate the dilution into 30 mL of LB for desired growth time (~30 min doubling time).
- Shake in falcon tubes at 37C until OD of 0.5-0.6
- Put cells in ice water bath for 30 min
- Wash Cells
- Spin down cells in rotor (max rpm)at 4C for 10 min
- Pour off supernate and re suspend cells in 30mL of cold water
- Repeat wash 2 times
- After final spin down, pour off the supernate and re suspend cells in ~300uL water
- Use 70uL allloquats for transformations
