Overnight Sanger Sequencing

Any Sample Types

Results by Noon Nextday

Sample Submission Guidelines

Plasmid Samples
  1. About 200 ng plasmid DNA is used in one Sanger sequencing reaction, the maximal volume of plasmid DNA used is 5 ul. We prefer plasmid with minimal concentration of 30 ng/ul.
  2. If the plasmid size is more than 20 kb, please double the amount of DNA.
PCR Samples, Purified or Unpurified
  1. We accept both purified and unpurified PCR samples. If samples are unpurified PCR, we will perform PCR purification before sequencing.
  2. For PCR product, we use about 10 ng DNA per kb, so please kindly provide PCR size information when making order.
E. coli colonies
  1. We accept agar plate, colony suspension, overnight culture.
  2. We will need to use one of these three methods to amplify DNA before sequencing. PCR is the first option we use, since this will allow us to deliver data the second day. RCA and plasmid miniprep will takes an extra day.
    • PCR: please provide amplicon size, forward and reverse primer.
    • Rolling Circle Amplification.
    • Plasmid miniprep: please provide antibiotics information.
  3. For bacterial suspension, place all samples in either tightly sealed 8-strip tubes or strip capped 96-well plate
Genomic DNA
  1. We have to perform PCR amplification before sequencing, please provide PCR primers and expected amplicon size
  2. Please send at least 10 ul DNA sample, with concentration ≥ 30 ng/ul
Sequencing Primer

We use 1 ul of 3 uM primer in 1 sequencing reaction, we take primer at 3 - 100 uM, please kindly provide primer concentration with your sample submission.

Free Primers

Available Common Primers (Complete List in Excel) || Search Online

Cost per sanger sequencing reaction is $5 if templates are plasmid DNA or purfied PCR. Extra cost is applied for other samples types (genomic DNA, colony, RCA, and unpurified PCR). Volume based discount is available, please email info@quintarabio.com for discount or quotation.

Premix Primer With DNA Sample

Premixed for success

Advantages of premixed samples:
  • Track samples easily
  • Get results faster
  • Enjoy low cost per reaction
  • Improve success rate significantly

Mixed in the same tube: Template + Primer

Unpremixed, DNA and Primer In Separate Tubes

Tube 1: Template     Tube 2: Primer Primer (5 pmol/μl or 5 μM), 10 μl. Or, use our free primer library.

Primer For Sequencing: Prefer Tm: 50-60C, GC content: 40-60%

E. Coli Colonies
Pick a colony and resuspend in diH2O or Tris Buffer

Two sets of tubes:

Tube 1: Colony Suspension     Tube 2: Primer

  1. Colonies need to grow for at least 16hrs at 37°C to reach good visible size
  2. Pick a single colony with sterile tip and resuspend in 30ul sterile diH2O or Tris buffer (10mM, ph8.0), 15 ul will be submitted to us, and the rest 15 ul will be kept by clients for future uses
  3. Prepare 2 separate tubes: 1 contains 15ul colony suspension, 1 contains 5ul primer at 5pmol/ul

How to Label Sample Tubes

Label the samples with your initials followed by numbers

8-strip PCR tube for samples containing template

Example: Samples from John Smith

Label on the cap as well as on the side of the tube

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Frederick, MD 21703