Sample Submission Information:

Our sequence analysis is performed on two 16 capillary ABI 3100 model DNA Sequencers. The sequencing reactions utilize a novel 3'-fluorescently-labeled dideoxynucleotide triphosphate mixture (BigDye Terminators from ABI) and Taq FS DNA polymerase in a thermal cycling protocol. The combination of these chemistries leads to a more accurate and sensitive analysis. A good quality DNA template can yield an average read length of 500-700 base pairs with >99% accuracy.


Chris Smith analyzes data from one of the ABI 3100 DNA sequencers










How To Ship

Pricing

Sample Submission Sheet

Sample Submission Guidelines

Template Preparation

Primer Design Information
 
 

How to ship:

Please overnight your samples, via FedEx or other express carrier.  In this case, samples will not need to be shipped on ice. Address shipments to the following address:
  Chris Smith
Marine DNA Sequencing Facility
MDIBL, Old Bar Harbor Rd.
Salisbury Cove, ME 04672Pricing: MDIBL/INBRE Investigators         $9/sequence

Outside Investigators                     $11/sequence

Sample Submission Guidelines: Provide primer and template premixed according to the following guidelines. The recipes are sufficient for two sequencing reactions, so if a technical failure occurs, we will not have to contact you for additional sample material.

l For DS plasmid DNA:

500-600 ng ds plasmid DNA template
2 µL 4µM primer
x µL sterile distilled water
24 µL total volume l For PCR products: Mix as above, except use 10 ng template for every 100 base pairs of PCR fragment length, up to 100 ng l For Cosmid, BACs and Phage Lambda DNA: Mix as above, except use 2000-3000 ng DNA template Template Preparation: Successful DNA sequencing results are highly dependent upon the quality of the DNA template supplied by the user. Careful quantitation of your DNA is crucial!

l Concentration of template should be based upon OD260 results

l Verify the concentration and check for overall quality by gel electrophoresis

l Provide the template in sterile distilled H2O (preferred) or 10mM Tris-HCl, pH 8.0, but not in TE buffer, as EDTA may interfere with the sequencing reaction

 Primer Design Information: Consider the following guidelines when designing primers for sequencing:

l 20 - 30 nucleotides in length

l Avoid runs of an identical nucleotide, particularly four or more Gs

l Keep the G/C content within 40-70%; the closer to 50%, the better

l A melting temperature of at least 50 C

l Avoid primers that can hybridize to form dimers

l Primers should ideally be located 30-40 base pairs upstream of region of interest