SOS Call to all experinced "Sequencers"

Jim jmeador at adnc.com
Mon Apr 1 09:44:34 EST 1996


Stephanie,

I would like to suggest that you consider sending them to a sequencing
service. Provided that the DNA is clean and free of a smeary background on
an agarose gel, you should be able to sequence all three with the end
primers and have both strands sequenced fully. That's 6 sequences in all,
and the reactions are running around $45-60 per reaction here in San
Diego. The 600bp fragment should sequence all the way if the primers work
well and the service you use knows what they are doing. 

If the reactions are $50/ apiece and you do 6 reactions, then it will cost
$300. A Sequenase kit is $200 and the 35S-ATP varies, but you can just
about be assured of spending more money doing it yourself even if you
don't pay yourself.

Good luck,
Jim


In article <199603281727.BAA03464 at relay3.jaring.my>, spyong at pc.jaring.my wrote:

Hello,,

The only sequencing I've ever done is through the text book. However, it
seems like now is the time to start sequencing. I have three PCR products
and need to sequence them before I work further only to find that they are
junk. Someone suggested end-labbeling, others say double strand sequencing,
some say cycle sequencing....some say stick to 'Sequanase' (whatever that
means). Do I have to subclone into M13?

Please,I know sequencing is not easy and may be take ages, but I just need
to know which would be to best method and time saving. My bands are
200bp,350bp and 600bps long.

Thanks a million.

My e-mail : spyong at pc.jaring.my

Cheers,
Stephanie



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