Radioactive probes by PCR

[Klaus Salger]

Daniel L. Burgess (Daniel.L.Burgess at um.cc.umich.edu) wrote:
: 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?
<snip>
: 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

Daniel,
why do you use random primers at all? Wouldn't it be smarter
to use just the PCR primers (just one for strand specific probes)?

  Klaus

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