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Heat inactivation/Gel purification or both?


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#1 laurequillo

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Posted 27 October 2009 - 08:34 AM

Hi there, I have a question regarding the heat inactivation of restriction enzymes and SAP. When I do a ligation normally I cut my plasmid, add SAP (if necessary) and then run and purify from a gel. My question is, should I heat inactivate the restriction enzyme and the SAP even if I purify from the gel? I never did, but reading old post in the forum I found that some people prefer to do it in that way. What do you think?

Thanks!
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#2 Vini

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Posted 28 October 2009 - 06:52 AM

Hi there, I have a question regarding the heat inactivation of restriction enzymes and SAP. When I do a ligation normally I cut my plasmid, add SAP (if necessary) and then run and purify from a gel. My question is, should I heat inactivate the restriction enzyme and the SAP even if I purify from the gel? I never did, but reading old post in the forum I found that some people prefer to do it in that way. What do you think?

Thanks!


hey, in our lab, we proceed w/o carrying out the heat inactivation and cloning works fine 4 us this way. i really don't think its necessary. i guess e'one likes to follow their lab practice, hence differing views. personally, i don't see the need to do it. :)

#3 laurequillo

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Posted 10 November 2009 - 06:50 AM

Good to know.
Thanks!
"He must be very ignorant for he answers every question he is asked" Voltaire

"This is SPARTA!"

"Im the goddamn batman"




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