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DNA SEQUENCING FAQS

Q: How much template do you need for sequencing?
A: The required template amount is dependent upon the template type (see below) but all template should be submitted at a minimum 6μl volume.

  • Plasmids should be submitted at a maximum of 100 ng/μl
  • PCR products should be submitted at 25ng/1μl for DNA 1 KB or less in length
  • Primers should be submitted at 3 pmol/ μl

Q: What is the turnaround time I can expect?
A: Samples submitted before 10 am will be available to customers within 48 hours. Please note the following times that we process samples.

  • Samples submitted for cycle sequencing after 10AM will be processed the following business day.

Q: What solution should my samples be resuspended in?
A: Final elution of DNA must be done in ddH20 rather than TE or the EDTA solution often provided with DNA isolation kits.

Q: Does sequence context affect my results?
A: Yes, Please let us know if your template is GC or AT rich (>70%) so that we can adjust the chemistry accordingly.

Q: My reactions keep failing…..why?
A: Reactions fail for a number of reasons. The most common failures are due to poor quality template, template too concentrated or primer too dilute. It is critical to accurately quantitate the template. Primers should be checked for Tm, GC content, and dimer formation as well as having a unique primer site on the template.

Q: Do you accept premixed (primer/template) samples?
A: Yes, please follow the conditions above.

Q: Do you accept primer-walking projects?
A: Yes, we are able to perform primer-walking projects within the MGC. We would do the initial sequencing runs based on information provided by the customer and then edit sequence, design primers and sequence again until we obtain complete coverage on both strands. The customer is then provided with the edited consensus sequence and the primer stocks. Please contact the MGC for more information.

To schedule an appointment or to discuss experimental design, send an emai to Andrew Kanostl, or call 409-777-0392.