In article <4asort$c6b at legba.synergy.net> grant@ writes:
>Subject: DNA concentrations in Sequenase X'ns ??
>Date: 15 Dec 1995 21:21:00 GMT
>Standard protocol of Sequencing reactions with unlabeled primer and labeled
>dATP using Sequenase use a minimum of 5 microgram/reaction.
>Has anyone tried using 2 micrograms or below ??
>Will increasing the amount of primer (to say 50 pmoles/reaction) help the reaction
>produce better banding.
>If someone has an idiot proof protocol on these lines, can you please post them
>in this newsgroup.
I routinely use between 0.5 and 1.0 ug of plasmid and 1 pmole of primer with
the standard Sequenase protocol and get great sequence.