problems propagating pBluescript - reaaaaaally weird, help! (Mar/03/2006 )
has anyone ever had problems propagating pBluescript? when i grow up bugs (DH5a) for maxiprepping they grow really well (in the presence of ampicillin), yet when i maxiprep them i get no yield. i know the problem isn't the maxi, because other plasmids i prep at the same time always come out with high yield... anyone have any ideas? thanks in advance.
-Nubbins-
QUOTE (Nubbins @ Mar 4 2006, 05:33 AM)
has anyone ever had problems propagating pBluescript? when i grow up bugs (DH5a) for maxiprepping they grow really well (in the presence of ampicillin), yet when i maxiprep them i get no yield. i know the problem isn't the maxi, because other plasmids i prep at the same time always come out with high yield... anyone have any ideas? thanks in advance.
I have made maxi of pBluescript and worked well always. It is weird since other plasmids work well. You may have contamination in your original plasmid, check it is really pBlue, also check if your ampicillin is ok (is not expired for example and it is the correct concentration ), also try other cells to transform. You migth do minis for all this test first. Hope it can help though it is really extrange for me
-virolo-