On Wed, 31 Mar 1999 10:03:53 +0200, Marcus Wernitz
<viacx at mlucom6.urz.uni-halle.de> wrote:
>In trying to save money we did TOPO cloning in half the given volume.
>Now we need twice as much One Shot cells.
>Who has a good idea to get selfmade One Shot Cells or who tried a
>transformation in other E.coli strains with good results?
>>Any comment welcomed!
i had a sort of similar situation using the TA cloning kit where i
required more competent cells, so we just grew about 10 ul of OneShot
cells up to 200 ml of log phase culture and put them to a procedure to
render them chemically competent before storage at -80C. these cells
weren't quite as transformation-efficient as the originals but were
plenty good enough to use. i can dig up the protocol if you like..
i am very interested in how you stretched out your TOPO kit and how
well that worked.. could you give more details?