Introduction
The CU Cancer Center DNA Sequencing & Analysis Service is a component of the newly amalgamated CU Cancer Center Molecular Pathology Shared Resource. This facility aims to provide reliable DNA sequencing data rapidly to its customers. This goal is accomplished using state-of-the-art capillary DNA sequencing instruments (ABI 3730s) with turn-around times of usually one business day.

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.

Contact this facility for current pricing. These services are priced very competitively for both academic and industrial researchers.

In addition to DNA Sequencing & Analysis services, we also provide:

• DNA profiling for authenticating human cell line identity
• Identification of mutation in genes involved in human cancer development and tumor responses to chemotherapy by the SNaPshot assay
• Multiplex Ligation-dependent Probe Amplification (MLPA) analysis for determining gene copy number
• Consulting and trouble-shooting for DNA sequencing, mutation detection, PCR, and gene cloning projects

Descriptions of Core Protocols and Acknowledgements for Publications

Use of the facility for services is with the understanding that the Molecular Pathology Shared Resource is acknowledged in research publications which utilize it. This is critical for future funding. 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