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“It was the best of times, it was the worst of times” (C. Dickens: A Tale of Two Cities) The Biomolecular Core Laboratory (BCL) provides essential services in molecular biology and genetics to the research and clinical staff at the Alfred I. duPont Hospital for Children and affiliates.
In 2004, the Biomedical Research Department was awarded a Center for Biomedical Research Excellence grant from the National Institutes of Health for the establishment of a new center for pediatric research. Funds were available for BCL to upgrade existing DNA sequencing and genotyping services and establish new high-throughput gene expression services. There were many challenges encountered when optimized assays were moved from the ABI Prism 377 DNA Sequencer to the ABI Prism 3130xl 16-capillary Genetic Analyzer.
Customers experienced difficulties when using sequencing protocols that were optimized for the slab gel system, on the new capillary system. The wide variety of genetic tests offered for molecular diagnostics had to be re-evaluated and optimized to fit the enhanced capabilities of the new platform. Even the chromatogram printouts looked different, and it took our customers (and ourselves) some time to adjust the settings to fit customers’ needs.
In this poster, we will outline the steps involved in updating sequencing technology from slab-gel to capillary electrophoreses and present how, through multiple rounds of trial and error, customers and core staff were able to re-evaluate existing assays and identify several important factors (DNA purity, BigDye dilution factor, buffer conditions, removal of dye blobs, software settings) that contribute to sequencing success. We will also report on a customer survey that was conducted to evaluate core performance, identify areas needing improvement, and highlight the strengths of our new programs and the steps taken to improve customer satisfaction, data quality, and cost effectiveness.