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Can transormations be incubated at less than 37 degrees after heat shock?


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4 replies to this topic

#1 KimWG

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Posted 28 November 2009 - 12:20 PM

Hi,

I came into the lab on a Saturday to set up some transformations from a ligation I ran overnight. The incubating shaker in our lab gets a lot of use so I was hoping to be able to sneak my guys in there after their heat shock. However, someone has something in the incubating shaker at 30 degrees (don't know who, or what). Can I incubate the transformations at this lower temperature, perhaps for a longer time, or should I just store my ligations until the incubator is free again? I'm trying to generate some results for a grant proposal due in a week, so I'm really under the gun here!

Thanks
Kim

#2 phage434

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Posted 28 November 2009 - 01:07 PM

There should be no problem in incubating at 30C for a bit longer. I would run the transformation efficiency control so that if I ran into problems I would be able to pinpoint this as a possible cause, but I would not anticipate a problem.

#3 Qundo12

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Posted 29 November 2009 - 09:23 AM

No problem for sure. I usually do that and colonies formed well ^^ (but I didn't make any comparison to evaluate the efficiency)

#4 KimWG

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Posted 29 November 2009 - 10:39 PM

No problem for sure. I usually do that and colonies formed well ^^ (but I didn't make any comparison to evaluate the efficiency)


Thanks all -- The shaking incubator was freed up by the time I needed it but it's good to know for future reference!

#5 tfitzwater

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Posted 30 November 2009 - 07:50 AM

When I transform temperature sensitive plasmids, after a 42C heat pulse I add SOC and incubate at 30C for 90 minutes, plate, and incubate the plates at 30C for about 6 hours longer than normal.




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