sequencing riddle...

David Grant dgrant at iastate.edu
Tue Apr 15 19:32:41 EST 1997


In article <5i4afv$9k1 at net.bio.net>, Hiranya Roychowdhury
<hroychow at NMSU.Edu> wrote:

> Hello All,
>         We have come across an interesting brain teaser. One of our
> colleagues has amplified a piece of DNA from a plant using a single primer
> (i.e. amplified region bordered by inverted repeats...). Now the teaser
> (may not be one for some of you): How can this piece of DNA be sequenced
> WITHOUT having to CLONE it? In other words, the investigator wants to use
> the same primers to sequence the pcr products directly after purifying
> them! This conundrum has occupied parts of our brains for the last week or
> so. I tend to think that this is impossible.... but then there may be some
> clever way which is eluding us...
>
if you're lucky, there is a single site for some RE asymetrically located
in the fragment - if so cut w/ RE, gel purify fragments individually and
sequence them

DG

David Grant   dgrant at iastate.edu
USDA-ARS and Iowa State University






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