pcr dna purification for sequencing

Chris LaRosa clarosa at biocomp.unl.edu
Fri Aug 4 09:08:02 EST 2000



Alex Plakantonakis wrote:
> 
> hi everyone.
> 
> how are you?
> 
> my question is:
> 
> what is the best method of purification of pcr products for sequencing?
> could you perhaps sketch the protocol briefly?
> 
> hopefully someone will the take the time to help me out by answering this
> question.
> 
Best is sometimes a subjective idea. Also it is important for you to
state what sequencing protocol and instruments you are using.  The best
method will depend on your procedure.  I spent a year doing sequencing
on a ABI 310 and in my opinion the Gibco Concert pcr purification
cartriges where cost and result effective.   However, recently the users
in that lab that I left have had good sucess using a sodium
acetate/alcohol precipitation.  
http://lamarck.unl.edu/research/systematics/Orti/GO-lab/protocols/MasterProtocol.html   
. Visit the site below to review the current lab protocols used in a
molecular systematics laboratory for ABI based sequencing.






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