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

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...

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Detalles Bibliográficos
Autores principales: Lennon, Niall J, Lintner, Robert E, Anderson, Scott, Alvarez, Pablo, Barry, Andrew, Brockman, William, Daza, Riza, Erlich, Rachel L, Giannoukos, Georgia, Green, Lisa, Hollinger, Andrew, Hoover, Cindi A, Jaffe, David B, Juhn, Frank, McCarthy, Danielle, Perrin, Danielle, Ponchner, Karen, Powers, Taryn L, Rizzolo, Kamran, Robbins, Dana, Ryan, Elizabeth, Russ, Carsten, Sparrow, Todd, Stalker, John, Steelman, Scott, Weiand, Michael, Zimmer, Andrew, Henn, Matthew R, Nusbaum, Chad, Nicol, Robert
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2872875/
https://www.ncbi.nlm.nih.gov/pubmed/20137071
http://dx.doi.org/10.1186/gb-2010-11-2-r15
Descripción
Sumario: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.

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