Double stranded dna Sequencing help
marcusj at helix.mgh.harvard.edu
Mon Aug 5 13:58:42 EST 1996
<Pine.A18.104.22.1680802132330.104918B-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 ?
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,
P.S. I have no connections with either Amersham or Oncor.
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