Recommendations

For best results, we suggest the following; (Also check our helpful hints page!!!!! )

Prepare the template DNA and primer in DI water...Do not use TE.
Primer must be at least 18-24 bp with a melting point temperature of 50oC higher.
The primer must be at a concentration of 3.2-10 pmol/ul.
Indicate type of DNA template being sequenced; ds, ss, or pcr DNA (include size)
For each reaction: 50-100 ng ssDNA, 0.5-1.0 ug cosmid DNA
0.5-1.0 ug dsDNA, 1-2 ug BAC, 2-3 ug bacterial genomic DNA
For successful direct genomic sequencing we strongly recommend a high quality genomic prep.
For recommended DNA isolation methods please contact us for details at This email address is being protected from spambots. You need JavaScript enabled to view it. . We recommend Mo Bio Laboratories for DNA isolation kits for the ease and quality of the preps as well as the low cost per sample.
Carefully check all templates for proper concentration and purity... LOW TEMPLATE CONCENTRATION, SALT,and or ALCOHOL contamination in preps will result in failure of sequencing reaction !!!
For sequencing a PCR and DS product:
100-200 bp 10-20 ng/rxn
200-500 bp 20-40 ng/rxn
500-1000 bp 30-80 ng/rxn
1000-2000 bp 50-100 ng/rxn
>2000 bp 100-200 ng/rxn
single-stranded 50-100 ng/rxn
double-stranded 200-500 ng/rxn
cosmid, BAC 0.5-1.0 ug/rxn
bacterial genomic DNA 2-3 ug/rxn
Adjust to a final volume of 10ul and label tubes w/ DNA concentration and date.