Q:protocol for cycle sequencing of cosmid templates

Armin Michel armi at stud.uni-sb.de
Tue Jun 17 03:01:38 EST 1997

Does anybody know of a reliable protocol for sequencing cosmids by cycle
sequencing? Of special interest for me are the amounts of DNA template
and primer to be used in the cycling reaction. I tried the sequencing
reaction with the standard template amounts for plasmid DNA (about 500
ng which yielded good results in our standard plasmid sequencing
reactions) and with significant higher template concentration (about 2
µg) but got no usable sequence using standard sequencing protocols
referring to the ABI 373/PE ABI Prism Cycle Sequencing manual. 

Every help is greatly appreciated.

Thank you in advance,
Armin Michel

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