University of Colorado DNA Sequencing & Analysis Core
Sample Guidelines Table | DNA Sequencing Services & Sample Guidelines | DNA Sequencing troubleshooting | Quantification of DNA using Mass Ladders | Human Cell Line Identification & Authentication Guideliness
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Introduction
The University of Colorado DNA Sequencing & Analysis Core Laboratory, now a service component of the Department of Pathology, aims to provide reliable DNA sequencing data rapidly to its customers in the Colorado Front Range. This goal is accomplished using state-of-the-art capillary DNA sequencing instruments (ABI 3730s) with turn-around times of usually one business day.

We will continue to operate in our current locations and provide all of our customers with reliable and rapid (overnight) turn-around of Sanger DNA sequencing results at market competitive prices. With the restructuring of the Core it has been possible to significantly reduce the prices of our sequencing services, which are now very competitive for all researchers (as low as $3.40/reaction). To see our competitive prices log in or download the pricing flyer.

Customers can supply the DNA as one of the following: plasmid DNA, DNA fragments (restriction or PCR products), lambda phage DNA, cosmid DNA, or BAC DNA. Please refer to the DNA Preparation Guidelines links on the left and the links at the top and left of this page to download our handbooks for DNA preparation and troubleshooting problematic results. The table below provides a summary of specific sample requirements to ensure optimal results.

We have an expansive menu of other available services. In addition to DNA sequencing, we can also perform:

    • DNA profiling for authenticating human cell line identity (we are a reference testing lab for the International Journal of Cancer);
    • Identification of ~70 mutation in genes involved in human cancer development and tumor responses to chemotherapy by a clinically validated SNaPshot assay;
    • Multiplex Ligation-dependent Probe Amplification (MLPA) analysis for determining gene copy number (amplifications and deletions), and
    • Various other fragment size determination assays.

Moreover, we continue to develop new services and expand others to meet our customers' needs utilizing capillary electrophoresis.

Furthermore, as we have done over the past 17 years of operation, we offer free consulting and knowledgeable and extensive trouble-shooting assistance on DNA sequencing, mutation detection, PCR, and gene cloning projects. To support these services, we have developed an extensive library of protocol manuals and trouble-shooting guides and handbooks, which are available on this website (see the links on the left and top for troubleshooting guidelines and our troubleshooting manual/handbook in the download section).

Should you have any questions, please do not hesitate to contact either our new manager Robby Shelton at 303-724-3166 (Robert.Shelton@ucdenver.edu) or the Core's director Christopher Korch at 303-724-3164 (Christopher.Korch@ucdenver.edu).


Descriptions of Core Protocols and Acknowledgements for Publications

Use of the facility for services is with the understanding that the UC DNA Sequencing & Analysis Core is acknowledged in research publications which utilize it. See this link for Acknowledgement and Service's Protocols for publications.

Guidelines for Submitting Samples
For premium reactions, DNAs and primers are submitted separately; whereas, for mid-level reactions, DNAs and primers are pre-mixed by the customer.

The premium service can be useful when a customer requests sequencing with one of the standard primers available through the Service , requires sequencing with one of our special sequencing chemistry mixtures for difficult templates, or wishes to sequence one template with multiple primers. In the last case, the customer uses less DNA per reaction and can retrieve the remaining DNA from the facility.

The Service's primers and special sequencing chemistry mixes are only available as a premium service.

Drop boxes at the University of Colorado Anschutz Medical Campus at 12801 East 17thAvenue.

  • 8th Floor of RC-1 South next to the elevators (picked up throughout the day)
  • 5th Floor of RC-2 next to the freight elevators (picked up twice per day)

Samples received after 4PM may not be processed until the next business day.

Optimal Concentration
Submit samples in
Reaction type
Volume
Plasmid
PCR Product &
Restriction Fragment***
BAC, Cosmid, & Phage
1.5mL microfuge tubes
Premium (DNA, Primer Separate)
DNA
10 μL, +5 μL per reaction**
100-200 ng/μL
10-15 ng/μL per kb
500 ng/μL
Primer
10 μL, +1 μL per reaction**
5 microMolar
5 microMolar
50-100 microMolar
Mid-Level (Pre-mixed DNA + Primer)
10 μL
750 ng DNA +
10 picomoles Primer
30 ng/kb DNA +
10 picomoles Primer
NA
96-well plates*
High throughput (HT)
(Pre-mixed DNA + Primer)
10 μL
200 ng/well DNA +
5 picomoles Primer
15 ng/kb DNA +
5 picomoles Primer
NA
 
DNA Profiling
Discuss project with the facility's personnel before submitting samples
 
Special Project

Look at the DNA Preparation Guidelines link on the left for specific requirements for your samples.

Additional instructions for HT:
*Contact the Service for details. 96-well plates must be compatible with Applied Biosystems 9700 Thermocyclers.
Please leave one well empty for pGEM control (H11 for 47 samples, H12 for >48 samples).

Additional instructions for Premium:
**If more than one reaction per template is requested, please add 5 μl for each additional reaction.
Submit a minimum of 10 μl of the primer (at 5 μMolar in water) for the first reaction and add 1 μl of primer for each additional reaction.
Note: Premium Plasmid template guidelines are for plasmids under 10kb.

Additional instructions for PCR & Restriction Frament templates
***Adjust the DNA amount in proportion to the fragment/product length

 

 

 

©1995-2012 University of Colorado Anschutz Medical Campus DNA Sequencing & Analysis Core