DNA Sequencing

The DNA Sequencing Facility provides: Automated Sequencing using capillary-based AB 3130 and AB 3730 DNA Analyzers. These instruments offer high throughput and sensitivity with a capability to handle 400 to 800 samples per day. These DNA Analyzers allow the Facility to have turnaround time of 1 to 2 days. Samples of single stranded, double stranded DNA, and PCR products, can be sequenced with reads >800 base pairs and a success rate of over 90%.

Sequencing requests are entered using our on-line dnalims web server at http://coreweb.icmb.utexas.edu/ (146.6.133.14). The samples are then brought to MBB 1.426 and put into the green "core prepared" or pink "user prepared"; racks in the refrigerator. An email will be sent when the samples are sequenced. The sequencing files can then be downloaded at coreweb. For a more in depth explanation, pricing and sample deadlins look at the information below or see Sequencing Handout, FAQ, or Troubleshooting.

Individual samples - Core or User Prepared Facility does cycle sequencing and cleanup.	$4.00/sample Deadline for samples 10 am
96-well Plate Samples - Core or User Prepared User submits 96 samples in a 96 well plate. Samples should go down (vertical) with sample 1 at A1 to sample 8 at H1, etc.	$300/plate Anytime

Core Prepared Information

A Core Prepared sample is where the cycle sequencing reaction and clean up is performed by the Core Facility and loaded on the sequencer. Individual samples should be brought in a 1.5 ml microfuge tube with a total volume of 12 µl of DNA and primer mix. Plasmid DNA should be 500 ng with the primer at 5-20 pmol. For PCR samples submit 5 ng/100 bp with 5-20 pmol of primer. Some primers are available free of charge. Plate samples should be in a 96 well pcr plate with the samples numbered down the plate (vertical). Samples should be brought to MBB 1.426 and placed in the cold box on the west wall. Core Prepared samples should be placed in the green rack. Facility Map
For more information see the FAQ or Sequencing Handout

User Prepared Information

User Prepared samples are where the customer does the cycle sequencing reaction and clean up. The Core Facility resuspends the sample in liquid and runs the sample on the sequencer. Individual samples should be brought in a 1.5 ml microfuge tube. Plate samples should be in a 96 well pcr plate with the samples numbered down the plate (vertical). Samples should be brought to MBB 1.426 and placed in the cold box on the west wall. User Prepared samples should be placed in the pink rack. Facility Map
For more information see the FAQ or Sequencing Handout.

Sample Submission Process

Sample requests are submitted electronically by using your browser and going to http://coreweb.icmb.utexas.edu (http://146.6.133.14). You then need to login or create a login. Login to enter sequencing requests. Use the link "Enter DNA Sequencing Requests" enter the number of samples, fill out the form and submit. For more information see the FAQ or dnaLIMS Instructions.

Data Retrievable Process

Sequencing data can be retrieved electronically by using your browser and going to http://coreweb.icmb.utexas.edu (http://146.6.133.14). After login in use the link "Download DNA Results" and you can select the order number/date to retrieve results. The results can be viewed in the browser window using the java applet, or you can download the results to view them with the appropriate application.
For more information see the FAQ or dnaLIMS Instructions.