Transforming with large AT rich inserts - (Jun/24/2005 )
I am using Invitrogen's multisite gateway cloning kit and am having troubles keeping my insert from recombining out! Sequencing shows that about 30bp of my inserts still exist at either end but the middle 2-5kb are gone. I have tried TOP10 and DH5a, both with recA1 but am looking for other competent cells that might be more accepting of the AT rich sequence.
SURE cells probably would be a better cloning choice for these kinds of problems.