Amersham kit problem resolved

Stephen Perry bafq5 at central.susx.ac.uk
Sun Oct 19 18:50:58 EST 1997

About a week ago I posted a request for information about problems
associated with Amersham Dye primer kits, partly in response to our own
difficulties and from reading of other's problems. I have since received
several emails with problems, including a gel image file (sorry, but I
was unable to read this file, so couldn't compare to our own). Both a
colleague and I have spoken to Amersham reps and discussed the problems.
No other reports of problems with this batch have been made to Amersham,
and our problems (short read length, poor signal) seemed to be specific
to our DNA, as the control DNA still sequenced perfectly. However, we
had not altered our DNA prepping protocols, so the poor performance
still looked like a kit problem. We managed to resolve this by
increasing the extension times on the PCR protocol to 2 minutes (from 30
sec), but the results were still variable, but usable. 
I have since spoken to an Amersham rep who has asked me to post this to
the group to clarify that there is no detectable problem with their kit,
and the issue is still being resolved. I am waiting to hear back about
whether their QC dept have found any difference in this batch at all,
which might explain why highly purified DNA sequences perfectly, why
less pure DNA does not, even though this level of purity has not been
problematic in the past.
Thank you to eveyone who wrote to me, sorry I have not replied to you
individually. I will keep you informed of any further developments. 

Steve Perry

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