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DH10BAC

baculovirus bluo-gal iptg DH10BAC transposition

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3 replies to this topic

#1 Guest_Imran_*

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Posted 09 October 2012 - 09:12 PM

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

#2 bob1

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Posted 10 October 2012 - 12:09 AM

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

#3 Curtis

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Posted 10 October 2012 - 01:19 AM

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

#4 bob1

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Posted 10 October 2012 - 12:31 PM

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





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