The WU Biology DNA Sequencing core facility has the ability to run your DNA sequencing samples and return the results to you by electronic means.

Facility Use:

To use the facility you need to prepare the DNA template for sequencing as well as prepare and run the cycle sequencing reaction yourself. Your samples will be run on our DNA sequencing machine, currently an ABI 3130xl.  For labs running very small numbers of samples (less than 20 a month) it is unlikely to be cost efficient to use the Biology Dept core facility. For these very low throughput users I recommend that you outsource the entire cycle-sequencing and analysis process. Users in this category have reported satisfactory results from the PNACL  at the medical school.

We only accept DNA samples that have been sequenced with Applied Biosystems BigDye DNA sequencing kit. However we do accept both BigDye version 1.1 and BigDye version 3.1.

Sample Preparation and Troubleshooting:

There is a lot of online information on how to prepare template for sequencing, how to run cycle sequencing reactions and how to clean and dry your samples (if you choose to do the clean-yup yourself).  I recommend you start with the BigDye manual available from Applied Biosystems (www.appliedbiosystems.com).  Once there, search for “BigDye terminator manual” under “product & technical literature” to get links to the ABI recommended protocols.

Another web site with help on DNA sequencing comes from the Washington University core facility at the Medical School, the Protein and Nucleic Acid Chemistry Laboratory (PNACL).  On the services page there is a very good PDF available that gives a lot of good advice on how to do DNA sequencing.

Sample Submission:

Once you have prepared samples, you can drop them of anytime in 112-south McDonnell Hall. Simply follow the instructions posted there on the freezer that is just to your left as you enter the room. The room is locked with the B94E key, this is a generic key available to any one in the department, simply complete the necessary key form and have it signed by your advisor (See anyone in the stock room for more information on department key policy).

All samples dropped off by 10:30 a.m. are run that same day and the results are usually posted on the Sequencing center server by the end of the workday. It is our goal is to get your data posted as soon as possible. We have over a 90% success rate of posting data by the end of day they were submitted. However, during particularly busy periods your data may not be posted until the next day. Samples are run on a first come first serve basis but we reserve the right to postpone the processing of large batches (> 48) of samples for up to 3 business days.

Data Retrieval:

To obtain your data use any mac in the department to connect to “Sequencer” from the network. If asked, connect to the server as “guest”, and navigate sequences>sequence data. Once there sequence data is organized by PI name and date.

Charges:

For samples supplied in individual tubes the charge is $9 per sample, however Biology department members receive a $4 per lane cost sharing that gives them a net cost of $5 per sample.

We offer a sequencing clean-up protocol in which we will do the post cycle sequencing clean-up for $1 per sample.

For more than 16 samples there are other services available.  See the medium throughput and high throughput sections of the web site for information on these services or contact Chris Shaffer for more details.