REQUIREMENTS FOR SEQUENCING

Your DNA template must be very clean for successful sequencing. Make sure you follow recommended methods of preparation and adjust your concentrations properly. Don't assume that because a particular kit or protocol for isolation works for another lab procedure, it will work with our system. It may not. See our Sample Preparation page for information on preparing template.

Resuspend your DNA template in very clean dH2O. The tris buffer that comes with the Qiagen kits will work if it does not contain EDTA. Do not use TE! The EDTA cheletes the magnesium ions required for our enzyme to work. Primers that are stocked in TE can be diluted with dH2O if the dilution factor is large(>5).

Do not use JM109 as a host strain. It makes excessive amounts of protein and it is difficult to get a preparation free of protein contamination.

Protein contamination and improper concentrations are the most frequent causes of inadequate sequence. In addition, improperly prepared sample can be detrimental to our capillary array. Please make every effort to give us high quality samples!

Qiagen publishes a very good template purification guide. It would serve you well to download and read it. Please ignore its advice on genomic preparation as we do not accept genomic preps.

Concentrations and Volumes:

Concentrations are important! Please don't submit samples that are not at the requested concentrations! There is much to do and stopping to dilute multiple samples with different concentrations takes up too much time.

Plasmids: 50-100 ng/ul and 4-5ul for every reaction.

PCR Products: Please see our Sequencing PCR Products Page for concentrations and volumes.

Where to send your samples:

For UAMS/Children's Hospital customers:

A valid UAMS account number is required. We will not process your samples without one. If you don't wish to use a UAMS account number, please provide a PO number. We are located in Biomedical Building I, room 504. Please submit samples after 10:00 am.

For non-UAMS customers:

A valid PO number is required. We will not process your samples without one. Please ship your samples to the address shown below. We highly recommend you use packing and seal your tubes with parafilm. Please ship your samples using the exact address shown below.

Allen Gies, DNA Sequencing Core Facility

Dept. of Microbiology and Immunology, Slot 511

Biomedical Building I, Room 504

4301 W. Markham

Little Rock, AR 72205

501-686-6927