PCR/sequencing

Steven Sullivan sullivan at gwis2.circ.gwu.edu
Sun Jun 1 04:19:12 EST 1997


Zhonglin Chai (zchai at COBRA.PATH.MONASH.EDU.AU) wrote:
: Mark Tucker (mtucke39 at IDT.NET) wrote:
: : does anyone have a one tube PCR/sequencing protocol that works well?  I
: : appreciate any help given.

: : mt

: I have been using Perkin Elmer's ABI PRISM Dye Terminator Cycle Sequencing
: Ready Reaction Kit wiht AmpliTaq Polymerase, FS. I works beautifully, and
: very easy to handle (just add your primer and template to a PCR tube and
: mix with 8ul of the terminator ready reaction mix, 25 cycles, and ethanol
: precipation, that's it). I routinely use it to sequence both PCR products
: and plasmid DNA. Email me if you need more details.

I agree that it works great.  It is, however, quite expensive ($675 for
100 rxns).  Also, we've had best success with column-purified reactions
(Centri-Sep), rather than EtOH-precipitates, and that runs into even more
money.  Has anyone tried similar kits from other manufacturers? 



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