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A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454

2010; BioMed Central; Volume: 11; Issue: 2 Linguagem: Inglês

10.1186/gb-2010-11-2-r15

1474-760X

Niall J. Lennon, Robert Lintner, Scott Anderson, Pablo Álvarez, Andrew J. Barry, William W. Brockman, Riza M. Daza, Rachel Erlich, Georgia Giannoukos, Lisa Green, Andrew Hollinger, Cindi A. Hoover, David B. Jaffe, Frank Juhn, Danielle McCarthy, Danielle Perrin, Karen Ponchner, Taryn L Powers, Kamran Rizzolo, Dana Robbins, Elizabeth M. Ryan, Carsten Russ, Todd Sparrow, John Stalker, Scott Steelman, Michael Weiand, Andrew Zimmer, Matthew R. Henn, Chad Nusbaum, Robert Nicol,

Biosensors and Analytical Detection

We present an automated, high throughput library construction process for 454 technology. Sample handling errors and cross-contamination are minimized via end-to-end barcoding of plasticware, along with molecular DNA barcoding of constructs. Automation-friendly magnetic bead-based size selection and cleanup steps have been devised, eliminating major bottlenecks and significant sources of error. Using this methodology, one technician can create 96 sequence-ready 454 libraries in 2 days, a dramatic improvement over the standard method.