3 replies to this topic

[Imran]

Hi all,

I have transformed some dh10bac cells with pfastbac vector and plating them on x-gal, bluo-gal, kan, tet, and gentamycin plates. all i am getting is white colonies (which is good), but i was wondering if transposition can be so good that i dont get any untransposed colonies? i tried plating Dh10bac straight on to the plates above (minus gentamycin) and am still getting white colonies. is it possible that either IPTG or bluo gal is not working?

any help would be really appreciated

Cheers,
Imran

[bob1]

Definitely could be that the IPTG is not working, it's not very stable even at -20.  Only lasts about a month.

[Curtis]

what is the difference between DH10B and DH10BAC. A derivative?

[bob1]

Forgot to add, it could be that the plasmid you are using doesn't have a B-gal marker on it too...