Sequencing high G + C DNA

Allen Gathman C741SCB at SEMOVM.SEMO.EDU
Tue May 2 11:34:56 EST 1995

In article <3o5dk4$rv3 at> Steven Goldberg <goldberg at> writes:
>I have been attempting to sequence DNA of high G + C content (ca. 72%)

(snip, snip)

We have similar problems.  Although we haven't yet figured it out, we
are about to try the suggestions in a paper titled "General method for
amplifying regions of very high G+C content" from NAR 21:2953-2954.

Basically, the authors say to use denaturation temp of 98 C, five min
combined annealing/elongation at 70 C, and to try reducing template
concetntration.  (Obviously, I'm assuming you're using a cycling
sequencing protocol here.)

We're about to try this; will let you know.  I'd appreciate hearing any
other suggestions.

|    Allen Gathman                |                                    |
|    Biology Department           |  Internet: C741SCB at SEMOVM.SEMO.EDU |
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