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 e-mail 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 product:
- 100-200 bp 1-3 ng DNA
- 200-500 bp 3-10 ng DNA
- 500-1000 bp 5-20 ng DNA
- 1000-2000 bp 10-40 ng DNA
  >2000 bp 40-100 ng DNA
Adjust to a final volume of 1-10ul and label tubes w/ DNA concentration and date.