CORE--DNA SEQUENCING FACILITY

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The Automated DNA Sequencing Facility is a merged Facility which provides investigators with state-of-the-art DNA sequencing and analysis of DNA polymorphisms. The Facility is used by 52 peer-reviewed funded investigators across 10 Research Programs from all three Divisions. Ninety-nine and one-half percent (99.5%) of the users have peer-reviewed funding. Richard Hardy, Ph.D. (Program Leader, Immunobiology) assumed direction of the Facility in 1993. Automated sequencing using an ABI Model 373 was set up in 1994. Model 377 sequencers were subsequently added, one in 1995, and another in 1997. A third 377 replaced the original 373 in 2002. Recently, two of the 377s have been replaced by a modern ABI 3100 capillary instrument, providing more reliable high-throughput sequencing. The Sequencing Facility provides Fox Chase Cancer Center (FCCC) investigators with computer-readable sequences of their DNA samples in a timely and cost effective manner, generating data more rapidly and of greater read-length compared to manual sequencing. Automated sequence analysis is used by investigators to verify DNA constructs, to identify mutations, and to determine the structure of newly cloned genes. Sequencing large genes (5-20 Kb) is much more tractable using automated sequence analysis, generating results in a few weeks, as compared to months using manual sequencing. The volume of usage of the Facility has continued to increase steadily over the past five years from 17,000 sequences in 1998 to over 28,000 in 2003 (64% increase). In addition to sequencing, analysis of DNA polymorphisms, important in genetic mapping studies, is also available in the Facility using "GeneScan" software on the 377. Equitable access to the Facility is assured by a Faculty Oversight Committee. The Facility was rated "Outstanding" at the last CCSG review.