Non-radioactive labelling of DNA

And'ys bionet andrew.bettany at bbsrc.ac.uk
Tue May 9 02:36:09 EST 2000


Hi,

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,

Andy

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
>p32
>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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