gateway cloning

| |Koen via methods%40net.bio.net (by k.vd.wetering from nki.nl)
Wed Jan 7 04:51:54 EST 2009


Hi,

we are currently using the Invitrogen Gateway system to make
expression vectors for our genes of interest. We do so by using
pDonr223 and our PCR amplified cDNA containing the recombination
sites. For some genes this works well but for others we have
difficulty in getting colonies containing the cDNA of interest. Our
problem does not seem to be due to the length of the cDNA as we have
succesfully made Entry clones containing cDNAs up to 5000 bp without
any problem. Any ideas about the reseans for the varying efficiency of
generating Entry vectors using the pDonr and BP-clonase 2 and the PCR
amplified fragment containing recombination sites?

Koen


More information about the Methods mailing list