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

ABSTRACT: 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.

SUBMITTER: Lennon NJ

PROVIDER: S-EPMC2872875 | biostudies-other | 2010

REPOSITORIES: biostudies-other

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

Lennon Niall J NJ Lintner Robert E RE Anderson Scott S Alvarez Pablo P Barry Andrew A Brockman William W Daza Riza R Erlich Rachel L RL Giannoukos Georgia G Green Lisa L Hollinger Andrew A Hoover Cindi A CA Jaffe David B DB Juhn Frank F McCarthy Danielle D Perrin Danielle D Ponchner Karen K Powers Taryn L TL Rizzolo Kamran K Robbins Dana D Ryan Elizabeth E Russ Carsten C Sparrow Todd T Stalker John J Steelman Scott S Weiand Michael M Zimmer Andrew A Henn Matthew R MR Nusbaum Chad C Nicol Robert R

Genome biology 20100205 2

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 dramati ...[more]

PMID: 20137071

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