PolyT sequencing

Galvin, Nora 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.  

Nora Galvin
Bristol-Myers Squibb Company




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