Best primers for ABI sequencing of Bluescript

RM Brownlie rmb10 at unix.york.ac.uk
Wed Aug 23 10:17:11 EST 1995


I have inserts cloned into the EcoRI/XhoI site of the Bluescript II SK 
vector and would like to know which are the best primers for Dye 
Terminator cycle sequencing using an ABI automated sequencer. Perkin and 
Elmer recommend the M13 forward and reverse primers but the reverse 
primer is a considerable distance from the EcoRI site. Has anyone had 
much success using the T7 or T3 standard primers or modifications of and 
did they modify any of the standard conditions? I would also like to know 
whether it is worth doing single strand rescue to obtain maximum read 
lengths?
Any information regarding achieving optimum results would be gratefully 
received.

Robert Brownlie




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