i just started working with cloning....my problem is that i did n't get any transformed colonies my vector is pcr2.1 of 4000bp, and my insert size is 1620bp, digested with Kpn1 and Bamh1. i checked the efficiency of my competent cell,its fine,checked the efficiency of ligase enzyme.its fine
Then i icreased the insert:vector ratio from 1:3,1:5, increased incubation..1hr to overnight. but no colonies....can any one help me in this problem?
Edited by priyak, 26 March 2009 - 07:41 AM.