2 replies to this topic

[laurequillo]

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!

[Vini]

laurequillo, on Oct 27 2009, 11:04 PM, said:

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.

[laurequillo]

Good to know.
Thanks!