AW: Amersham ET terminators for 3700

Verena Bohle vbohle at oncoscreen.com
Mon Oct 13 03:07:51 EST 2003


Hello everyone, hello Phillip,

if you reduce the amount of BigDye to 1/16th, do you reduce the amount of D=
NA in the same proportion?

Has anyone tried to reduce the extension time using BigDye? What did you no=
tice then?

Thank you for any comments
Verena Bohle



-----Urspr=FCngliche Nachricht-----
Von:=09Phillip San Miguel [SMTP:pmiguel at purdue.edu]
Gesendet am:=09Samstag, 27. September 2003 19:02
An:=09autoseq at net.bio.net
Betreff:=09Re: Amersham ET terminators for 3700

Paul,

=09I have, but never tried below 1/2 reactions. Amersham's polymerase/chemi=
stry
allows much quicker thermal cycling (extensions are only 1min instead of 4 =
min).
But I think their polymerase is not as thermal stable. So while BD runs fin=
e for
99 cycles--at least in the presence of 5% DMSO, I think the Amersham chemis=
try
will not.

=09Also BD v3.1 under version 2.0 of data collection in our hands just give=
s
better results than Amersham's chemistry.

=09If you are looking to save money, why not drop to 1/16th reactions with =
the Big
Dye--that works fine.

Phillip SanMiguel
Purdue Genomics Core Facility

Paul Shinn wrote:
>    Does anyone use Amersham ET Terminators for sequencing plasmid on the
> 3700?  We've been using BigDye for years but I can save a considerable
> amount if I can switch to the ET Terminators.  I need to dilute them to
> 1/6rxn to save any money (I do 1/8 BD now).  Can you dilute it this low
> for the 3700?  Are you happy with the quality?
>
> =09=09=09=09Thanks, Paul
>
>
> ---
> Paul Shinn                   pshinn at neomorph.salk.edu
> SIGnAL                       http://signal.salk.edu
> Sequencing Coordinator       http://genome.salk.edu
> (858) 453-4100 ext 1796
>

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