DNA sequencing

Recommendations from EUROFINS

DNASizeConcentrationVolume
Plasmid DNA-50-100 ng/µl15 µL
Purified PCR products150-300 bp2 ng/µl15 µl
Purified PCR products300-1000 bp5 ng/µl15 µl
Purified PCR products1000-3000 bp10 ng/µl15 µl
Unpurified PCR products150-300 bp4 ng/µl15 µl
Unpurified PCR products300-1000 bp10 ng/µl15 µl
Unpurified PCR products1000-3000 bp20 ng/µl15 µl

Quantify the template concentration via agarose gel or a photometer to ensure accurate results.

  1. Put barcode stickers on all tubes
  2. Add 15 µl purified DNA with either of the concentrations given in the table above
  3. Add 2 µl of primer with a concentration of 10 pmol/µl (10 µM)
  4. The total volume of your premixed sample should be 17 µL

Recommendations from GATC

Sending your samples the right way Sample requirements: Take 5 µl template DNA with either of following concentration:

  • Purified plasmid DNA: 80 - 100 ng/µl
  • Purified PCR products:
    • 150-300 bp: 2 ng/µl
    • 300-1000 bp: 12 ng/µl
    • 1000 bp - : 25 ng/µl

Add 2.5 µl of primer with a concentration of 10 µM (10 pmol/µl) Please send total sample amount of 10 µl in 1,5 ml tubes To ensure the best possible quality of sequence data, the total volume of the sample should not be less than 10 µl. We recommend measuring the DNA concentration on an agarose gel.