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.
<--SNIP-->
>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.
>
>Alan.
<--SNIP-->

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.

George

P.S. We use the 377s.

-- 
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