Random primed labelling trouble

Paul Ulanch Pulanch at bioch.tamu.edu
Thu Sep 26 14:04:19 EST 1996


In article <52ambt$ln6 at merkurius.lu.se>,
   Fredrik Kamme <fkhero at biogen.wblab.lu.se> wrote:
>Hi everybody,
>I've got a random primed labelling trouble:
>I'm using a DNA labelling kit with random decamers (MBI-Fermentas) that
>works fine on Qiaquick gel eluted fragments. However, when I try
>labelling a 220bp DNA piece (also gel eluted plasmid insert) I get no
>labelling. Tried it twice, but no success. Is the DNA piece too short 
and
>re-anneals too quick? Can there be a contamination in the gel-eluted 
DNA?
>
>Many thanks in advance, Freddy

The primers in the kit are random decamers?  The problem may be that the 
primers are too big!  We use random hexamers from Pharmacia (I have no 
relations with that company) in our 'home-ade" kit and they work great. 
 Try smaller primers, maybe this will work with such a sshort probe.  
Good luck!

Paul "Goose" Ulanch

Dept. Biochem/Biophys
Texas A&M University
College Station, TX   77843-2128
(409) 845-0832
PULANCH at bioch.tamu.edu



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