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Competent Cells - (Sep/28/2009 )

Does anyone know if in the process of purifying competent cells, should the culture be at a specific optical density for the best results?

-Biochick99-

I didnt get you ...........

-macroman-

Yes! Check out the original paper by Hanahan 1983.

-bob1-

Biochick99 on Sep 28 2009, 10:41 PM said:

Does anyone know if in the process of purifying competent cells, should the culture be at a specific optical density for the best results?

yes, the OD600 must be at 0.5 which is the log phase of bacterila growth.

-banou-

Biochick99 on Sep 28 2009, 01:41 PM said:

Does anyone know if in the process of purifying competent cells, should the culture be at a specific optical density for the best results?


They do need to be at a specific OD but that will be different depending on what bacterium you're making competent. They need to be in log phase. If it's E coli that will be ~0.5 - my way of determining this is when you can only vaguely see the lab light thru the culture!

-microgirl-

0.3 to 0.6 is fine.

sometimes i just took an overnight, diluted 100 fold in fresh LB and let it grow for 2-3 hours.

Never let me down.

-hanming86-