Help for Double Digestion

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
joacocell
Garter
Garter
Posts: 2
Joined: Thu Apr 06, 2006 9:29 am

Help for Double Digestion

Post by joacocell » Fri Apr 21, 2006 2:59 pm

Hi everybody,

I want to make a construction (protein fusion) with the vector ECFP-C1.

I have linearize my vector with EcoR-I and XbaI,,,besides I have done the double digestion as well for 2 hrs because more than 2 hr my enzimes cut all my vector,,,my insert (PCR product) has cut with both enzimes as well overnight. vector and Inser were purified by gel. I have done the ligation overnight ,,the transformation,,however when I analyze my minipreps I haven't found any clone with my insert. I would like to say that I have run controls for my ligation and my transfromations and they worked fine.

I don't know what's the problem,,

Thanks

Post Reply

Who is online

Users browsing this forum: No registered users and 9 guests