3' end labelling with tritiated thymidine...

Rory O'Brien via methods%40net.bio.net (by rory.obrien from otago.ac.nz)
Mon Oct 12 15:32:48 EST 2009

Cheers Duncan,

The question came from a student here who wants to measure uptake of an
oligo into bacterial cells and the effect of a drug on same. The thought was
to label the oligo and measure via scintillation counter. I had thought we
could end tail the oligo with tritiated thymidine but she's now wary of
sticking a whole bunch of nucleotides onto the end of the oligo and the
effect that might have on her uptake experiment. I doubt we'd get a strong
enough signal with just end labeling and I'd rather avoid P32 as we're not
really set up for it here.

Could we do something non-isotopic with digoxygenin do you think? Maybe a
dot blot of lysed bugs and semi-quantitative chemiluminescent detection?

- Rory

Historians believe that in newspost 
<mailman.70.1255061417.1133.methods from net.bio.net> on Fri, 9 Oct 2009, 
Rory O'Brien <rory.obrien from stonebow.otago.ac.nz> penned the following 
literary masterpiece:
>Could anyone maybe suggest which thymidine product might be most
>to this application? Detection to be via scintillation counter; one for the
>old-school molecular biologists maybe ;-)

Highest specific activity would be best so either of the last two. 
Whether they are the least expensive is another matter. If you are 
simply using it to check the incorporation even the first one should do. 
You will just see lower counts.

I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.
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