ABI's Taq FS

George F. Mayhew mayhew at genetics.wisc.edu
Mon Oct 16 11:30:51 EST 1995

In article <lyons-1610951436480001 at cyclid.demon.co.uk>,
lyons at cyclid.demon.co.uk (Alan Lyons) wrote:

>In article <hardy-1410951419130001 at mighty.facs.fccc.edu>,
>hardy at mighty.fccc.edu (Richard R. Hardy) wrote:
>> In article <45k6c3$ie1 at nntp3.u.washington.edu>, jason2 at u.washington.edu
>> (Jason Seto) wrote:
>> >Has anyone used the Taq FS from ABI and found they have not had to purify
>> >the sequenced product prior to electrophoresis on a 3373/377?
>> >
>> >Does the EtOH ppt step work, or do you lose the first few bases?
>> >or do people still use the spin columns to purify off the excess dyes,
>> >regardless of ABI's claims that you don't need this step with their new 
>> >enzyme/kits?
>> We have been getting good results with the new Taq FS kit from ABI; we
>> compared sequences from the old kit (w/spin column) to that with new kit
>> (EtOH ppt) and got equivalent or (in most cases) better sequences on an
>> ABI 377.
>I would agree, we get equivalent or (in most cases) better sequences on an
>ABI 373, and you certainly do not need to phenol extract or use spin columns.

We've stuck with the "column" purification for the terminator kits,
because it's quicker when you use a 96-well plate containing the resin
(G-50).  Single samples have been ETOH ppt and we've seen no loss of
signal over what we get with the spin-plates.  We've also tried loading
directly without purification.  The terminator kits still seem to produce
too much background (T-blobs, flashes, smears, etc.), but the primer kits
(esp. when done 1/2 volume) give very little recognizable background
beyond the primer peak.

Hope this helps.


P.S. We use the 377s.

"People are DNA's way of making more DNA."(Edward O. Wilson, 1975)
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