Non-radioactive labelling of DNA
andrew.bettany at bbsrc.ac.uk
Tue May 9 02:36:09 EST 2000
My first reaction to Chris' reply was one of violent disagreement. however,
on reflection I find myself sort of agreeing with him. For whatever reason I
never seemed to get efficiently labelled DIG probes using random-primer
labelling - BUT I have got really good results using PCR DIG labelling to
make probes for Southerns and following the Roche protocols (can supply
references if you're really interested).
The Northerns have proved a little more difficult, but there appears to be
light at the end of the tunnel there too. Using DIG-labelled riboprobes
worked really well in my hands, but I could never strip the probe off
afterwards!! We noe seem to be having success using PCR DIG-labelled DNA
probes for northerns.
Certainly, in my experience of Southern blots, the non-radioactive
technologies compete very favourably with their radioactive counterparts.
Hope this helps,
Chris LaRosa wrote in message <3916DD28.2109D09A at biocomp.unl.edu>...
>"D. Durnford" wrote:
>> I would like to limit the use of 32P in my lab and I am shopping for
>> non-radioactive labelling methods (random prime). I am primarily
>I think you are ill advised to try non-radioactive probing if you have
>labelled probing set up in your lab. In general I have observed
>nonradiactive labelling never works as well for northerns and southerns
>compared to good old p32.
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