sequencing

Bill Melchior, NCTR/FDA wmelchior at NTBTOX.NCTR.FDA.GOV
Tue May 12 07:55:17 EST 1992


In responding to the following comment about a sequencing problem:

>>Our gels occasionally show all four 
>>nucleotides lighting up in each lane across a run.

Bruce Roe says:

>Sequenase is famous for "template associate compression" you describe.

I assume that Bruce's comment is about the "strong pause" phenomenon, in which
secondary structure causes the polymerase to pause, leading to bands in all
four lanes at one or a few short regions; he suggests several ways to over-
come this problem.

I had interpreted the original query as something different: bands in all
lanes in all positions, so the entire film looks like the photo of a picket
fence.  This latter problem has to do with template purity.

The appropriate solution depends on what the problem really is.
________________________________________________________________________________
The opinions stated are mine, not those of NCTR or its sponsoring organizations.

Bill Melchior                                ||   "You have lawyers the way
National Center for Toxicological Research   ||    other people have mice."
Jefferson, AR  72079                         ||
(501) 543-7206                               ||   -Kenneth Duncan, English
                                             ||    Health & Safety Executive,
wmelchior at ntdoc.nctnet.gov                   ||    to US regulators



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