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. © 2005 Yang et al; licensee BioMed Central Ltd.
CITATION STYLE
Yang, G. S., Stott, J. M., Smailus, D., Barber, S. A., Balasundaram, M., Marra, M. A., & Holt, R. A. (2005). High-throughput sequencing: A failure mode analysis. BMC Genomics, 6. https://doi.org/10.1186/1471-2164-6-2
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