IGEM:Melbourne/2008/Protocols/competent cells

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  • yield 5ml ~ 100 aliquots of 50┬ÁL
  • need another overnight culture (5ml)

Procedure

1) Grow 130ml LB culture from the 5ml grew previously to OD600 of NO MORE than 0.45

2) Pour cultures into sterile 50ml falcon tubes

3) Pellet cells at 4 degree, 3000rpm for 5 min

4) Discard supernatant

5) Resuspend pellet in total volume of 50ml ice-cold TFB1

6) Chill on ice for 5 min

7) Pellet cells at 4 degree, 3000rpm for 5 min

8) Discard supernatant

9) Resuspend pellet in 5 ml of ice-cold TFB2

10) Chill on ice for 15 min

11) Aliquot on dry ice, then snap-freeze, then store at -80 degree

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