Double stranded dna Sequencing help

Jeremy Marcus marcusj at helix.mgh.harvard.edu
Mon Aug 5 13:58:42 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. I know this can be caused because of 
: secondary structures and so on, but I would like to know if somebody can 
: pass me on, let's say, the latest version of the best method for 
: sequencing doubled stranded DNA. I use the Pharmacia kit, does anyone 
: uses something else besides what they provide ?
: thanks,
: Mozart

I've been comparing Amersham (USB) Sequenase and Oncor (Appligene)
Fidelity sequencing systems.  The verdict: they are both excellent; the
Sequenase protocol takes about an hour less than Fidelity, but Fidelity
resolves secondary structures more reliably.  Also, Fidelity _requires_ a
glycerol-tolerant gel.  It's optional with Sequenase.  

Hope this helps,
Jeremy

P.S. I have no connections with either Amersham or Oncor.



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