Biotinylation of plasmid DNA

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Fri Apr 3 22:37:40 EST 1998


In article <3524cee4.17766106 at news.task.gda.pl>, wasowicz at moskit.art.olsztyn.pl wrote:
>jsg at mole.bio.cam.ac.uk (J.S Good) wrote:
>
>>Hi
>>
>>How do I attach biotin to plasmid DNA and to linear fragments of dsDNA and
>>ssDNA? 

>You can try Photobiotin from Vector. It is working quite nicely. It is
>coming with the protocol. You will need a  mercury vapour lamp. They
>offer the lamp also if you cannot find one at your lab. This is quite
>convenient and universal way of labeling proteins and nucleic acids
>with biotin. Contact me if you need more information.

Yes, this is very simple and convenient. Two notes: 

1. Regular halogen lamp (>300 W) works as well. In any case, to prevent your 
sample from heating, keep it on ice AND put a layer of ice-cold water between 
the lamp and the sample (I used one of those huge Petri dishes). 

2. Don't label too much. I once labeled to the point that most of DNA became
insoluble (it was with 11 carbons photobiotin; maybe does not happen with 9, I 
don't know).

        - Dima






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