Google Sign in options
Remember me
This is not recommended for shared computers

Sign in anonymously
Don't add me to the active users list

Privacy Policy
Sign In
Create Account

Javascript Disabled Detected

You currently have javascript disabled. Several functions may not work. Please re-enable javascript to access full functionality.

0

Heat inactivation/Gel purification or both?

Started by laurequillo, Oct 27 2009 08:34 AM

Please log in to reply

2 replies to this topic

#1 laurequillo

The Goddamn Batman!

Active Members
265 posts

1
Neutral

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!

"He must be very ignorant for he answers every question he is asked" Voltaire

"This is SPARTA!"

"I´m the goddamn batman"

Back to top

#2 Vini

Veteran

Active Members
178 posts

1
Neutral

Posted 28 October 2009 - 06:52 AM

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.