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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