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Home > Services > Core Sequencing
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Core Sequencing
Group Leader (Lucinda Fulton)
- Mapping
The GSC provides mapping services to all investigators. The GSC has the ability to generate maps using agarose gel fingerprinting methods. Fingerprints are assembled and the resulting data can be compared to sequencing information to provide a complete genomic view.
- Whole genome Sequencing
The GSC provides large scale, high throughput DNA sequencing services to all investigators. Researchers can submit genomic DNA for plasmid or fosmid library construction or BAC clones for end sequencing. The resulting subclones are sequenced in an automated high-throughput barcoded pipeline. The resulting samples are loaded on an ABI 3730 capillary sequencer to produce longer high quality reads. The sequencing data is then retrieved via a private ftp site.
- Clone Sequencing
The GSC provides large scale, high throughput DNA sequencing services to all investigators. Researchers can submit individual clones (BACs, cosmids, or fosmids) in glycerol stocks and the GSC will purify clone DNA for library construction. Plasmid libraries are constructed and used to produce sequence data from the submitted clone. Samples are loaded on an ABI 3730 capillary sequencer to produce longer high quality reads. The subsequent sequence data is assembled with phrap and the contigs as well as the sequence data is made available via a FTP private site.
- Sequence Refinement
The GSC offers sequence refinement services for both whole genome and clone based projects. This is process is accomplished using an initial automated directed pipeline called prefinishing and a more manual directed pipeline call finishing. Projects can be processed through only the automated pipeline or both the automated and manual process. Finished sequence data is refined to an error rate less than 1:10,000 bases.
- EST, cDNA Sequencing
EST stands for Expressed Sequence Tag, and is usually a short stretch of nucleic acid (usually less than 500 bp in length) that has been derived from a gene expressed in some cell or tissue being studied. "ESTs provide researchers with a quick and inexpensive route for discovering new genes, for obtaining data on gene expression and regulation, and for constructing genome maps. The GSC offers EST (cDNA) library construction and DNA sequencing in an automated high-throughput pipeline using the ABI 3730 capillary sequencer. The sequence data can also be analyzed in the GSC analysis pipeline which includes submission to dbEST.
- Individual subclone sequencing
The Core Facility at the GSC provides large scale, high throughput DNA sequencing services to all investigators. Researchers can submit individual subclones in 96 or 384 well trays. The GSC can then sequence the clones using either standard vector-based sequencing primers or primers chosen by the researcher. Samples are loaded on an ABI 3730 capillary sequencer to produce longer high quality reads. The sequencing data is then retrieved via a private ftp site.
- Individual amplification sequencing
The Core Facility at the GSC provides large scale, high throughput DNA sequencing services to all investigators. Researchers can submit PCR products in 96 well trays. The GSC can then purify the products by Exonuclease/SAP-treatment and the products sequenced using the appropriate primers (also submitted by the researcher in 96 well plates). Samples are loaded on an ABI 3730 capillary sequencer to produce longer high quality reads. The sequencing data is then retrieved via a private ftp site.
- Required Documents for Sample Submission
We have recently been advised by the University's administration that we should require a copy of the IRB approval letter. Therefore starting immediately all samples that are human or associated with human DNA or tissue must be accompanied by a copy of the applicable Washington University IRB approval letter. We have altered our project sheet and sample submission sheet to request this information. Since collaborators often have multiple ongoing projects, the applicable IRB approval letter must be included with each sample submitted to the GSC for any human samples or samples associated with human DNA or tissue. If you have any questions or need help determining if your samples require IRB approval please contact Lucinda Fulton at the GSC or the Human Subjects Research Protection Office. Thank your for your cooperation in this matter.
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