Radio-labelling EthBr stained DNA

cjs at canctr.mc.duke.edu cjs at canctr.mc.duke.edu
Tue Feb 11 13:41:35 EST 1992


In article <1992Feb10.002954.11436 at minyos.xx.rmit.oz.au>, s860780 at minyos.xx.rmit.oz.au (Clinton Grant) writes:
>	Does anybody know whether DNA taken from a stained gel (and GeneCleaned)
>is OK to label up with P32 (using an oligonucleotide labelling kit)? Does the 
>EthBr interfere with the labelling reaction?
>
>			Thanks in advance. 

If you use Seaplaque Agarose (FMC) you can actually label the DNA without
Genecleaning it....we do this all the time and get results which are
indiscernable from DNA which is not in LMP Agarose. This is with EthBr and
all....The probes also hybridize well, no detectable differences there either.

Just a warning, not all LMP Agaroses are the same, for example, none of this
works if you use FMC's NuSieve Agarose....I believe that they even specify in
their manual that Seaplque is compatible with labelling reactions. They
don't say anything about the EthBr, however, it does not appear to interfere.

	Hope this helps-
		Caryl



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