IUBio

Thermo Sequenase

Rick Rick
Mon Dec 16 10:13:05 EST 1996


Sue at net.bio.net,sb328 at columbia.edu (Susan A Baumes) writes:

>Does anyone have any experience using Amersham's Thermo Sequenase along
>with the energy primers that they have?  We're thinking of trying them,
>but would like some feed back first.
>
>Right now we're running ABI's Amplitaq FS dye terminator, ready reaction
>mix.  So if anyone has a direct comparison that would be great.

We've been using TS and ET primers for quite some time now and we're
extremely happy with both.  Switching to these reagents for general M13
sequencing immediately added about 100 bp to our average read length,
due to increased signal-to-noise and improved the basecalling as a
result of the more uniform peak height.

Comparison to the ABI Taq FS terminator kit is apples & oranges.  You
still see much more accurate basecalling with dye-primer chemistry
because even with an F->Y enzyme, all of the dye-terminators are not
equally incorporated.  On the flip side, the terminator chemistry will
resolve artifacts (compressions, stops) that the dye-primer chemistry
cannot.  For de novo sequencing, a good mix of both chemistries is
still preferred.

Hope this helps!

Regards,
Rick



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