automated sequencing on ALFexpress

Stephk at niv.ac.za Stephk at niv.ac.za
Thu Jun 12 15:04:12 EST 1997


Hi there,

We are using the ALF automated sequencer from Pharmacia for more then
a year now and we have encountered various problems with the machine.
The biggest problem though are the random mutations that we are
getting frequently which are not supposed to be there. 
Various people at our institute are sequencing different viruses (HIV,
Flu, measles, HHV8 etc) using different methods (such as different
cycle sequencing kits from Pharmacia or Amersham, the AutoRead Kits
from Pharmacia or the AutoLoad Kits from Pharmacia for sequencing of
PCR products...). 
All of us are getting deletions and insertions and random point
mutations where we know there shouldn't be any (confirmed by manual
sequencing etc.).

Our question is whether anybody has ever experienced anything like
that using the Pharmacia ALF sequencer??? 
We would really like to know whether this problem only occurs on our
individual machine and with our reactions or whether it is a common
problem.
Thanks....

Stephanie Kreis
National Institute for Virology
Private Bag X4
Sandringhma 2131
South Africa
Email: stephk at niv.ac.za





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