Competent cells
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* incubate in ice for 20min | * incubate in ice for 20min | ||
* centrifuge for 1min | * centrifuge for 1min | ||
| - | * | + | * aspirate |
* add 1/10 volume (50uL) of cold 100mM CaCl2 | * add 1/10 volume (50uL) of cold 100mM CaCl2 | ||
* vortex | * vortex | ||
Latest revision as of 18:02, 28 June 2006
Competent Cells
Preparation: DH5az1 overnight
- 50 time dilution of o/n: 50uL o/n, 2.5mL LB broth
- incubate until OD reaches 0.4(measure OD with spectrophotometer)
- transfer 500uL diluted o/n to centrifuge tube
- add 1/2 volume (250uL) of cold 100mM CaCl2
- vortex
- incubate in ice for 20min
- centrifuge for 1min
- aspirate
- add 1/10 volume (50uL) of cold 100mM CaCl2
- vortex
- incubate on ice for 1h
- add 15% of glycerol of the final volume
- ready for transformation or store in -80C freezer
