Q: purified primers for sequencing??????
bernard at elsie.nci.nih.gov
Tue Apr 22 00:50:09 EST 1997
In article <landryp.861641720 at alize.ERE.UMontreal.CA>,
landryp at ERE.UMontreal.CA says...
>I am just about to send a couple of samples to a sequencing service, and
>I noticed that they require "purified primers". I checked the primers
>that I have (Operon) and It is specified on the label "unpurified".
>What does the purification process implies? Also, what is wrong with the
>use of unpurified primers for sequencing?
>Thanks in advance,
Well, there's "purified" and "purified"...
I also habitually buy my primers from Operon and in their description
unpurified = deprotected and gel purified, whilst
purified = HPLC purified
I believe in these days of "cutting our own throat" oligo
prices some companies skip the gel filtration step and leave it all
up to you. Maybe you can use very crude oligos for some applications
(I've never done it) but gel purified oligos have been fine for me for
PCR (Taq) and sequencing (Sequenase). Some methodologies may require
the full HPLC purification. I have very little experience with automatic
sequencing but coworkers use the same grade oligo in our ABI machine
with good results.
If cost cutting is a prime motive then try a test run with the
[I have no affiliation with Operon except as a satisfied customer]
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)
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