Maximum insert size in a pGEM Teasy vector - (Jun/12/2005 )
has anybody got experience with the maximum size of the insert in a standard cloning procedure in a pGEM Teasy vector(2 kb) and the following transformation into JM109 high efficient competent cells (those provided with the pGEM Teasy kit). A colleague told me that due to the fact that pGEM Teasy is a high copy plasmid, the maximum insert size is something around 4kb - otherwise the bacteria simply will die off. I'm a bit sceptically but I haven't got any experience and I also couldn't find anything in the manual provided with the kit. My aim is it to subclone a 8kb insert into the 2.6kb pGEM Teasy vector. What is your feeling-will it work?
Thank you very much for your help!
Has nobody got an idea?
Thanks for your help!
Don't know if I can help, I think the biggest I've ever gone is around 5kB into pGEM-T easy. It went in OK although efficiency wasn't fantastic. Given that 4 or 5Kb works I would give 8Kb a try, I'd be interested to hear back how it goes. I suspect that you will get a drop in efficiency but if you only need one clone it is worth a go.