ABI sequencing primer question
Eric C. Anderson
anderson at pharmdec.wustl.edu
Tue Jun 27 17:07:17 EST 1995
<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.
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.
"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
st. louis, mo 63110
for a comprehensive list of bike related web pages... http://pharmdec.wustl.edu/~anderson/anderson.html
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