Haynes Lab:Notebook/Engineering PC-TFs/2013/02/05: Difference between revisions
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== | ==2/5/13== | ||
* | <font size=3>'''Bacterial transformation'''</font> | ||
* Added total volume (10.0 μL) to 50 μL chemically competent cells (e.g., BL21) in a 2.0 mL tube. | |||
* Incubated on ice for 2 min., heat shock at 42°C for exactly 45 sec., and placed on ice. | |||
* Added 800 μL sterile SOC medium. | |||
* Grow with shaking at 37°C for 30 min.(''Taped tubes to the shaker rack.'') | |||
* Pelleted the cells at top speed in a microcentrifuge for 3 min. at room temperature. | |||
* Discarded the supernatant. Resuspended the cells in 100 μL LB + antibiotic. | |||
* Plate cells on pre-warmed LB agar + antibiotic. (''I plated 100 μL of the cells. Then put beads on plates and shook.) Protocol didn't specify.'') Grow overnight at 37°C. | |||
* Check results tomorrow. | |||
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Latest revision as of 22:26, 26 September 2017
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2/5/13Bacterial transformation
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