Problems for cloning a gene

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

Moderators: Leonid, amiradm, BioTeam

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

Problems for cloning a gene

Post by joacocell » Thu Apr 06, 2006 9:57 am

I want to to insert a gene of 1.2 Kb to a vector in the EcoRI and XbaI sites.

This gene was amplified by PCR and purified by gel. The PCR product was digested in 5' with EcoRI and 3' with XbaI overnight and purified by gel. The ligation was performed overnight and the transformation was performed according to the subcloning Efficiency DH5-alpha. However,I couldn't get any colony in my plates.

I dodn't know if I should start again since the beginning the cloning.

Any recommendations?.

Thanks

Post Reply

Who is online

Users browsing this forum: No registered users and 4 guests