I would like to know how many labs are using kits from Amersham with ThermoSequenase II for sequencing on ABI machines. How do the results compare with BigDye?
Thank you.
Ales
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Ales Merta merta@volny.cz
Ales Merta, Jizni II 22/774, 14100 Prague 4,
Czech Republic tel./fax (+420)2-72766622 mobil: 0602/724568
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Tom,
We often sequence directly lambda phage DNA, without problems. It was not
so before, the big difference came with the ThermoSequenase kits and labeled
dideoxynucleotides. The template has to be in greater amount than usual
(i.e. more than what we use for sequencing PCR fragments or plasmids),
usually around 1-5 micrograms per 20-25 ul reaction. We take care to get
rid of the RNA by digestion with RNase, followed by Sephadex G-50 removal of
the ribonucleotides and other small undigested pieces of RNA.
Regards,
wagner yotov, Univ. of Montreal