Radioactive probes by PCR

Daniel L. Burgess Daniel.L.Burgess at um.cc.umich.edu
Wed Dec 28 11:50:27 EST 1994


In article <asheri-251294092545 at louis.ls.huji.ac.il>, asheri at vms.huji.ac.il
(Ella) wrote:
> 
> Does anyone have a good protocol for labelling PCR fragments with 32-P and
> using them as probes for Southern hybridization?
> 
> Thanks in advance for any help you can give.
> 
> Ella
> 
> Dept. of Genetics, Hebrew University of Jerusalem
> email: asheri at vms.huji.ac.il

One thing we've been doing to improve incorporation of label during
random primed labelling of smallish (150-500 bp) PCR products is to 
add a bit of both PCR primers (that were used to amplify the product) 
to the reaction, in ADDITION to the random oligo mix.  This skews the 
average size of the labelled product in your favor and presumably
also speeds up the reaction rate. 

Daniel.L.Burgess at um.cc.umich.edu
PhD-2-B (T - 10 months, and counting)
Dept. of Human Genetics
 
Positional cloning...it's not the kill, it's the thrill of the chase.



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