To submit samples to the MBCF, first complete a sequencing request.
- Standard Reaction/Data Collection: samples should be submitted in the following form:
DNA samples that are submitted for standard ‘reaction/data collection’ service must be accurately quantified and prepared. Samples submitted with inaccurate concentrations and/or with contaminants present will result in poor sequence results or a failed reaction, which will still be billed to the user.
- For plasmid DNA <12kb - provide 20 µL of DNA at a concentration of 40 ng/µL in water. For PCR products 300-600bp - provide 20 µL of DNA at a concentration of 10 ng/µL in water.
- For a custom primer, please provide 10 µL at a 10 µM concentration and include the Tm of the primer.
- For template/primer mix please provide a 20ul volume of template at 40ng/ul and primer at 1uM final concentration
- For PCR fragment DNA- amplicons must be purified/treated prior to DNA sequencing. Please inquire for details and/or protocols. If you would like to submit PCR reaction, we can treat/prepare your sample for sequencing by purification or ExoSap treatment. Please inquire about cost and sample submission.
- The following standard primers are provided: M13F(-20), M13R,T7, T7 Term, SP6, T3, CMV-F, PGEX-F, PGEX-R, BGH-R, Lucif Ctrm Rev, GFP-R, pNTR1A-F, pNTR1A-R, PQE30-F, PQE-Tri-F, PQE-Tri-R, E-GFP-R, and V5 Ctrm Rev.
After processing the results will be available to you from the webpage. Please login and click on “View your sequencing results”.
The sequence will be repeated at no extra cost if there is an operator error or machine malfunction. Please provide accurately quantified, high quality DNA for successful results.
Automated sequencing is very sensitive to template and primer preparation.
It is essential that the templates be free from ethanol, salts and other contaminants, as these greatly affect the quality of the sequencing results.
NOTE: WE WILL NOT REPEAT SAMPLES AT NO CHARGE IF THE DNA IS OF BAD QUALITY OR INACCURATE CONCENTRATION.
Here are a couple programs available to view chromatogram files (.ab1):
Chromas Lite – from Technelysium
Finch TV – free download from Geospiza
Chromatogram Explorer – from DNAbaser
Human Identity Testing, Cell Line Authentication, and Mycoplasma Testing:
- Please provide isolated/purified genomic DNA in a minimum volume of 15ul and a minimum concentration of 30ng/ul.
- For ID and CLA testing only- a cell pellet can be submitted and we can perform the isolation for you. Standard DNA isolation rates apply.
Barbara Davis Center
Anschutz Medical Campus
1775 Aurora Court, 4th floor, Room 4207
Molecular Biology contact personnel:
Mark Farrell, Core Manager