Overnight Sanger Sequencing

Any Sample Types

Results by Noon Next Day

Sample Submission Guidelines

Plasmid Samples

Quantity and Concentration Requirements:

  1. A standard Sanger sequencing reaction requires approximately 200 ng of plasmid DNA.
  2. The minimum volume for a plasmid DNA submission is 10 µL, with a minimum concentration of 30 ng/µL.
  3. For plasmids exceeding 20 kb in size, the DNA quantity should be doubled to ensure optimal sequencing results.

PCR Samples, Purified and Unpurified

Sample Types:

We accept both purified and unpurified PCR samples. In the case of unpurified PCR samples, we will conduct a PCR purification process prior to sequencing.

Quantity Guidelines:

  1. Approximately 10 ng of DNA per kb is required for PCR products.
  2. Please provide the PCR product size information at the time of order submission.

E. coli Colonies

Acceptable Formats:

We accept E. coli samples in the form of agar plates, colony suspensions, or overnight cultures.

DNA Amplification Methods:

  1. To prepare for sequencing, we employ one of the following DNA amplification methods:
    1. PCR (Preferred): This method allows for next-day data delivery. Please provide the amplicon size, as well as forward and reverse primers.
    2. Rolling Circle Amplification (RCA).
    3. Plasmid Miniprep: Include antibiotic information for the sample.
  2. Note: RCA and plasmid miniprep methods may extend the data delivery timeline by an additional day.

Sample Packaging for Bacterial Suspensions:

Ensure all bacterial suspension samples are placed in either tightly sealed 8-strip tubes or in a strip-capped 96-well plate.

Genomic DNA

  1. PCR amplification is necessary prior to sequencing. Please provide the PCR primers and the expected amplicon size.
  2. A minimum of 10 µL of DNA sample is required, with a concentration of at least 30 ng/µL.
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 $6 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 a sterile tip and resuspend in 30ul sterile diH2O or Tris buffer (10mM, ph8.0), 15 ul will be submitted to us, and the remaining 15 ul will be kept by the 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

MA Site

625 Mt Auburn St, Suite 105
Cambridge, MA 02138

CA Site

3563 Investment Blvd, Suite 2
Hayward, CA 94545

CT Site

231 Farmington Ave
Farmington, CT 06032

MD Site

5350 Partners Ct, Suite C
Frederick, MD 21703