plasmid purification from mammalian cells? - (Nov/16/2006 )
this may be a silly question!!!!!!!
i have inherited a transiently transfected hek cell line and want to check the plasmid has everything it should. i have been told what it has but the source is dubious!!!
is there a method out there whereby i could purify a plasmid from transiently transfected HEK cells??
would it work the phenol-chloroform way ??
Do you mean a stable transfected line or a cell that has an episomal vector. A transiently transfected cell will loose the plasmid over time. To check for the plasmid, the best thing to do is to do a genomic DNA prep and then use PCR with sequences specific for the plasmid.