DNA Sequencing Team
High-Throughput Sequencing Unit

The primary mission of the High-Throughput Sequencing and Genotyping Unit is to facilitate the unraveling of genetic information across the large and diverse collection of animals, plants and microbes that are the research focus of many of our faculty. The unit prepares libraries and generates both purified DNA templates and sequence data using the Sanger, pyrosequencing (Roche/454 GS-FLX), and Illumina/Solexa Genomy Analyzer (coming soon) systems. These resources are also critically important to the operations of its sister units, Bioinformatics and Functional Genomics. In addition, the facility provides DNA services for investigators requiring core DNA sequencing, fragment analysis, and custom oliognucleotide synthesis. All of the Unit’s services include consultation and assistance in all aspects of a project. Researchers are encouraged to discuss their projects with facility personnel who would be glad to provide information and quotes for their projects.

The High-Throughput Sequencing team provides a vast array of services for high-throughput sequencing projects. The unit offers both conventional Sanger sequencing and the next generation sequencing technology utilizing the Roche/454 GS-FLX and Illumina/Solex Genome Analyzer. The unit prepares custom library construction for EST (primary and normalized) and BAC shotgun library sequencing. For the Sanger dideoxy-termination sequencing, we can take a project from the beginning stages of plating bacteria, colony picking the library, then inoculating and culturing the bacteria, using the QpixII. Duplicate glycerol stocks are provided for customers. The plasmid preps used for sequencing are purified with the Qiagen 9600 BioRobots or the cost-effective Heat Lysis methods. Conditions for each project are optimized to produce the best sequencing data. Primers for sequencing are designed and made in house, and the Robbins Hydra is utilized for precise reaction set-ups. Cycling conditions are optimized on the MJ Research DNA tetrads. Big Dye terminator chemistry is used for Sanger sequencing reactions and all sequencing is performed on the ABI 3730XL capillary systems. All 384- and 96-well format plates are labeled with a barcode, and a high throughput laboratory information management system (HTLIMS) is used to track sample flow.

The ultra high-throughput sequencing service takes advantage of next generation sequencing technologies by the Roche/454 GS-FLX and Illumina/Solexa 1G Genome Analyzer systems. Pyrosequencing, using the GS-FLX has been available since summer, 2007 and delivery of the Illumina/Solex Genome Analyzer is expected by spring, 2008. Please contact Dr. Ryan Kim (wrkim@uiuc.edu) at 217-244-3480 or Chris Wright (clwright@uiuc.edu) at 217-333-4372 to discuss ways the staff can be of assistance in achieving your project goal and providing a quote for your project.

High-Throughput Sequencing and Genotyping Unit
Ryan Kim, Ph.D - Director
340 Edward R. Madigan Laboratory, 1201 W. Gregory Drive, Urbana, IL 61801
Phone: (217) 244-3480      FAX: (217) 265-5066      Email: wrkim@uiuc.edu