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Competent Cells


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#1 Biochick99

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Posted 28 September 2009 - 12:41 PM

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

#2 macroman

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Posted 28 September 2009 - 01:08 PM

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

#3 bob1

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Posted 28 September 2009 - 03:49 PM

Yes! Check out the original paper by Hanahan 1983.

#4 banou

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Posted 29 September 2009 - 05:43 AM

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.

#5 microgirl

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Posted 02 October 2009 - 08:12 AM

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!

#6 hanming86

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Posted 05 October 2009 - 08:34 PM

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.

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