ANDREI.POPOV at bbsrc.ac.uk (Andrei Popov) wrote:
>>Hi, Daniel and John:
>>>>The reference for "Direct Transfer of Plasmid DNA from Yeast to E. coli
>>by Electroporation": Nucleic Acids Research, Vol. 20, No. 4, 917
>>>>People who have tried it in my department got no luck. Hope it will work
>>for you. Please let us know if it works.
>>>>Qian Ouyang (qo22 at andrew.cmu.edu)
>>I also tried it a couple of times with different plasmids without any luck.
>The electroporation method works but is *EXTREMELY* strain-dependent !
I used SEY6210 and SEY6211 and it worked perfectly. I also tried it with
HF7c and some of our mutant strains with no luck.
One little trick always worked for me: I transformed plasmid into the actual yeast strain. If that worked, the direct transfer from yeast to E.coli worked as well. It seems that not the E.coli are the problem (JM109, DH5alpha and TG1 gave identical results) but the yeast strain one uses.
Bert Popping http://www.dur.ac.uk/~dbl1bp
Dept. Biol. Sci. http://www.dur.ac.uk/~dbl0www
University of Durham http://www.dur.ac.uk/
Durham, DH1 3LE