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PubMed · 15631628

High-throughput sequencing: a failure mode analysis.

George S Yang·Jeffery M Stott·Duane Smailus·Sarah A Barber·Miruna Balasundaram·Marco A Marra·Robert A Holt
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Abstract

BACKGROUND: Basic manufacturing principles are becoming increasingly important in high-throughput sequencing facilities where there is a constant drive to increase quality, increase efficiency, and decrease operating costs. While high-throughput centres report failure rates typically on the order of 10%, the causes of sporadic sequencing failures are seldom analyzed in detail and have not, in the past, been formally reported. RESULTS: Here we report the results of a failure mode analysis of our production sequencing facility based on detailed evaluation of 9,216 ESTs generated from two cDNA libraries. Two categories of failures are described; process-related failures (failures due to equipment or sample handling) and template-related failures (failures that are revealed by close inspection of electropherograms and are likely due to properties of the template DNA sequence itself). CONCLUSIONS: Preventative action based on a detailed understanding of failure modes is likely to improve the performance of other production sequencing pipelines.

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AutomationBiotechnologyChromosome MappingComputational BiologyDNA PrimersExpressed Sequence TagsGene Expression ProfilingGene LibraryModels, StatisticalPlasmidsPopulusSequence Analysis, DNASoftwareJournal Article

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George S Yang, Jeffery M Stott, Duane Smailus, Sarah A Barber, Miruna Balasundaram, Marco A Marra, Robert A Holt. 2005-01-04. High-throughput sequencing: a failure mode analysis.. https://doi.org/10.1186/1471-2164-6-2

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