Fluorescent sequencing of high MW dsDNA
krishnan at BORCIM.WUSTL.EDU
krishnan at BORCIM.WUSTL.EDU
Mon Mar 28 08:23:44 EST 1994
Hi folks!
To my reading, most of the fluorescent sequencing seems to be with single-
stranded DNA template and comparatively less with small plasmid templates,
or PCR fragments. What is the netland experience on sequencing double-
stranded templates of varying sizes (say range of 5-25 kb)? Dugaiczyk
et al (1992) NAR 20:6421-6422 reported that linearizing cosmid sized templates
improved accuracy. Has this been the general experience too for ds DNA
or is this required only when dealing with complex (=cosmid sized large)
templates? Also, could somebody tell me whether using dye-primer or
dye-terminator chemistries makes a difference in the length and/or accuracy of
read obtained, particularly when directly sequencing ds DNA?
I shall be glad to post a summary.
Much appreciated.
Raja
More information about the Methods
mailing list