Akaline denaturation of the DNA prior to adding it to the reaction will
increase the read length considerablely in gc rich regions .Use a x5
denaturing mix of 1.0M NaoH +1mM EDTA.Incubate at 37 degrees for 30
mins. Then neutralise with sodium acetate pH4.8 ( 1/10 the total volume
of the reaction) mix .Then precipitate with ethanol on dry ice ,wash
with 70%, dry and resuspend in the required vol.
so usually working from a miniprep it would be
40ul mimiprep
10 ul x5 denaturant
5ul sodium acetate
this would give enough for about 4-6 reactions depending on the
conc.Hope this helps.
Jim hughes