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Condition for Transformation - heat shock, "A novice"

broth time tubes

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#1 CAT

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Posted 25 October 2011 - 11:35 AM

Hi,

I am confused at reading different protocols for heat shock. Anyone please give a clue. Thanks! Posted Image

For the same strain (say XL-blue), I have found:
  

1. one writes NZY+ broth, while another one writes SOC.  

2. And for the time at 42C, One said 45 seconds, the other said 30 seconds.

3. Then for the tubes,   falcon 2059  polypropylene tube, VS  eppendorf vial

Edited by CAT, 25 October 2011 - 11:36 AM.


#2 bob1

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Posted 25 October 2011 - 06:22 PM

These methods are all based on work done by Douglas Hanahan published  in 1983.  You should read his paper to find out why...

In general most of these sorts of things don't matter too much, but can sometimes lower the transformation efficiency drastically.  The exact conditions you use should be optimised based on what you are using!

#3 CAT

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Posted 26 October 2011 - 05:51 AM

Thanks, bob1. This paper helps a lot. Great!Posted Image





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