Galvin#m#_Nora.WFD_Mail_Server at msmail.bms.com
Mon Nov 9 13:40:37 EST 1998
I have had success in sequencing through polyA regions using a polyT primer as
suggested in ABI's "Chemistry Guide" (published May, 1995). They suggest a
T25 primer with A, G, or C at the 3' end. I use T25VA, T25VG, T25VC and T25VT
to create a better anchor. Sometimes the reaction works with an equimolar mix
of the primers, which saves on reagents if it works. Other times I need to
try all 4 separately in order to get a clean sequence.
The "Chemistry Guide" is a great reference for troubleshooting sequencing.
Its main drawback is that it was published just before TaqFS was introduced,
and the chromatograms are from sequencing done with the old enzyme.
Bristol-Myers Squibb Company
More information about the Autoseq