Double stranded dna Sequencing help

Vlad Svetlov vvsvetlov at utmem1.utmem.edu
Sun Aug 4 20:16:16 EST 1996


In article
<Pine.A32.3.91.960802132330.104918B-100000 at spider.uspnet.usp.br>,
mdamarin at usp.br (Mozart de Azevedo Marins) wrote:

> Hi there,
>               I have to sequence three clones that I will use to express 
> in Drosophila. I am having some problems with the sequencing due to too 
> much stacking of the bands. 

> thanks,
> Mozart

I've been using fmol PCR sequencing kit from Promega and Genomyx PCR kit
to sequence ds plasmids - both work well and got rid of some compressions
that I've been seeing in my clones before. Both kits allow use of
alpha-S35 for direct incorporation and reliably read 500 bases from single
reaction (although Genomyx claims up to 750 my gels do not look nice at
such long runs due to poor heat dissipation). Personally I find PCR
sequencing easier to set up and less sensitive to sequence peculiarities. 

Best wishes, live long and prosper,
Yours, 
Salieri.



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