Subcloning and Mutations - (Oct/22/2008 )
we have pretty strange mutation problems with subcloning of a 2 kB human gene from a Teasy clone to pLenti.
By sequencing the pLenti-construct we found 1-3 mutations in the gene, which we haven't found in the Teasy clone.
Does this mean, that the polymerase of our competent cells is creating mutations in the vector? Have you seen this problem? Any ideas?
Thanks a lot!
You have to use a special competent cell line for lentiviral vectors as they recombine very easily. I use the Stbl3 competent cells from invitrogen. I think this might solve your problem.