Probe labelling by PCR

mikem at s-and-s.com mikem at s-and-s.com
Wed Nov 22 11:01:12 EST 1995


>   Thomas Urbig <Thomas at hastingslab.harvard.edu> writes:
>  Has onybody experience labeling gene probes by PCR and can provide 
>  reaction conditions for this. It should also be possible to make single 
>  stranded probes. If so, do you use a single stranded PCR product as 
>  template or is a standard (double strand) PCR product as template fine. 
>  Is the lower concentration of the 32P nucleotide a problem or do I have 
>  to add cold nucleotide? 
>  Any advice is welcome, thanks in advance
>    
>                 Thomas.
>  
>  
>  
>>>>
One can label oligonucleotides and use them as primers.  Also, one can label
the entire length of the PCR product.  It really depends on the sensitivity 
that you require for your method.  Schleicher and Schuell has the Rad
Free System which utilizes psoralen-biotin to label oligos and DNA in a
non-enzymatic, yet specific manner.  Additionally, exonuclease III can be
used to create single stranded probes of defined length.  For information
on any of these, respond to techserv at s-and-s.com or call
1-800-245-4024 and '2' for Technical Services.



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