pcr dna purification for sequencing

Biopolymer Facility biopolymer at sc3101.med.buffalo.edu
Fri Aug 4 13:20:06 EST 2000


>>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.
>>
>>thank you very much:)
>>
>>alex

One additional note regarding the Exo I/SAP protocol: this will work well
if you have optimized your PCR and are seeing only one band on a gel. If
you find you have multiple products on a gel, then of course you will need
to cut out your band of interest and then purify using a gel extraction
method in order to get good sequencing results. Qiagen and Millipore, for
example, have good commercial kits for gel extraction.

Michelle






>
>Hi Alex,
>I work in a DNA sequencing core facility and I find that some of the best
>results come from people who clean up their PCR reactions using an
>exonuclease/shrimp alkaline phosphatase protocol. I don't have the protocol
>in front of me but I believe it's relatively simple.
>
>Michelle
>
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>_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/
>Biopolymer Facility
>Roswell Park Cancer Institute
>Buffalo, NY   14263
>Phone: (716) 845-8032
>Fax: (716) 845-7621
>Email: biopolymer at sc3101.med.buffalo.edu
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Biopolymer Facility
Roswell Park Cancer Institute
Buffalo, NY   14263
Phone: (716) 845-8032
Fax: (716) 845-7621
Email: biopolymer at sc3101.med.buffalo.edu
_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/



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