2 replies to this topic

[ppc]

 cloning_problem.doc   24.5K   92 downloads

[aliaksandra]

For how long you incubate your ligation reaction? If I were you I would do the ligation at 4 degrees overnight. When I have an insert larger than 2kb, I always use these conditions.

[snoopyx]

Check your ligation by ligating a.) CIP treated and b.) untreated cut vector. So you can test if your ligation works and if your CIP treatment is OK. BTW I treat my vector with CIP usually just for one hour, it's more then enough for my protocol.

Edited by snoopyx, 02 April 2010 - 04:04 PM.