Does anyone have some words of wisdom for us. We are trying to
subclone a 10kB genomic fragment from a cosmid clone into a
plasmid. However, we are experiencing alot of difficulties. One
problem is that even though the DNA fragment is gel purified some
cosmid DNA must end up in our ligation mix and preferentially get
transformed. All clones we end up with appear not to be ligated into
the vector we are using, bit recircularized cosmid. Ideally we want a
vector with 2 antibiotic resistance genes and MCS (Sac1 site if
poss.). Does one exist? Anyother ideas to overcome our problems?
Andy Cannons
