DNA Sample Requirements

For the best results, please provide the clean DNA, (free of contaminants: salt, polysaccharides, RNA etc.), suspended in dd water (avoid 1 x TE buffer). (Some mini-prep kits do not ensure cycle-sequencing grade DNA, especially if the yield is low). The PCR products should be a single band on the gel and free of primers and dNTPs. This may be achieved by purifying the PCR products with the spin columns or equivalent methods. 

Please provide the accurate DNA concentration. The RNA or other contaminants in a DNA sample may yield artificially high concentration if measured spectroscopically (260/280 ratio). Check the sample on agarose gel. 

The amount and concentration of DNA sample required for sequencing:

Template Type DNA Concentration Total Volume
Plasmid 60 ng / µl 10 µl
PCR(500bp-1kb) 2-3 ng / µl 10 µl
PCR(>1kb) 4-10 ng / µl 10 µl
Cosmid 0.5-1 µg / µl 20 µl
BAC 0.5-1 µg / µl 20 µl
Genomic DNA 2-3 µg / µl 20 µl

Primers: No charge for Universal primers.
Custom primers: Please provide 10 µl primer solution at 10 pmol / µl, along with template, separately.  

Note: We encourage the use of primer design software.  For better DNA sequencing results, the following parameters help in creating custom primers:

  • 18-22 base long with 50%-60% G-C content
  • annealing temperature of 50 to 65 degree centigrade
  • absence of dimerization capability
  • absence of significant hairpin formation (>3 bp)
  • lack of secondary priming sites
  • low specific binding at the 3' end (to avoid mispriming)

* Not all primers will produce successful sequences.

DNA Sequencing Order Form: Please download Sequencing Order form, fill it and email your sequencing order

Results {Sequence & chromatogram Files}: The data will be e-mailed next day