Hi all. I have tried cloning a potential toxic gene about 1.2 kbp into E.coli with T7 promoter. After two months, I am still not successful. I am now plannig to do cloning into pUC18 vector before subcloning into few other vectors with different promoters. I have limited available restriction sites so that I don't have to redesign PCR primers. Is it feasible to do blunt-end cloning with Phusion polymerase into non-dephosphorylated vectors? Will I only be getting empty vectors after transformations?
Thank you. Matt _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
