Competent cells

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Line 10: Line 10:
* incubate in ice for 20min
* incubate in ice for 20min
* centrifuge for 1min
* centrifuge for 1min
-
* suck away supernatant
+
* 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

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