Origami and other folding strains
kaj.stenberg at helsinki.fi.invalid
kaj.stenberg at helsinki.fi.invalid
Mon Mar 19 01:49:08 EST 2001
"Michael L. Sullivan" <mlsulliv at facstaff.wisc.edu> wrote:
> Thanks for the input. In answer to your final question, no I haven't
> checked the few transformants for activity. I wasn't optimitic the few I
> did recover would have activity, since the numbers suggest there would be a
> strong selection against it. I suspected that the few transformants I was
> recovering had probably lost the ability to express an active protein. I
> waited to try the pLysS experiment first. Of course now I will go back and
> check it out.
Always check for activity/expression, after all, you only need one colony!
An anecdote:
I once tried to blunt-end ligate my gene of interest. Sorry to say, I
didn't get any colonies. Repeating with high volume and some 50
transformation plates I got two colonies. I tested them. One had the
insert the wrong way, one had it the right way!
_Sometimes_ things actually bounce your way in the lab! (It later turned
out that the home made competent cells were horribly bad).
--
Kaj
More information about the Methods
mailing list