Are transiently transfected DNA methylated in mammalian cells? - (May/25/2006 )
I would like hear from the experts here if transiently transfeced plasmid DNA to cells ( in the form of episome, no replication sequences within) are methylated overtime for the duration of its existance? Thanks.
I have had not much experience with transient transfections, I would have to say though, because it's foreign DNA, the cell can tell this and I would put my money on the fact that it would be methylated over time. Even more so if it's not replicated which is a means of passive demethylation.
Thats what I think too. Any suggestion how to prove that this is indeed the case? What are the best approach to do so. Would methylation sensitive RE good choice to get start with? Thanks in advance.
certainly, and then a probe with your vector on a southern would confirm this.