ABI sequencing primer question

[Eric C. Anderson]

In article
<Pine.A32.3.91j.950626095717.41060A-100000 at homer25.u.washington.edu>,
Peter Myler <mylerpj at u.washington.edu> wrote:
> We have had similar experiences here.  Primers with a lot of extra 5' 
> sequence generally do not make the best sequencing primers.  I would 
> suggest making new sequencing primers (they are very inexpensive, 
> nowadays), if more than 5-6 bp don't match at the 5' end.

peter,

thanks for the help.  something that i decided to try was to increase the
concentration of primers to make up for the fact that only 2/3 of the
primer was actually binding to the template.  it worked on the shorter of
the two primers but not the longer one.  unforturnately i will have a
tough time talking this lab into buying new primers, regardless of cost. 
they actually wanted to try sequencing this template with a 50-mer primer
that had a 15bp overhang.  oh well.

eric

-- 
"i don't know what caffeine does for you, but i'm pretty sure that without it your head caves in."

eric c. anderson                                 anderson at pharmdec.wustl.edu
dept. of molecular bio. and pharm.               (314)362-3963 (lab)
washington univ. school of medicine              (314)362-7058 (FAX)
660 s. euclid box 8103                           
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