Method
A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454
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* Corresponding author: Robert Nicol nicol@broadinstitute.org
1 Genome Sequencing Platform, Broad Institute of MIT and Harvard, 320 Charles St., Cambridge, MA 02141, USA
2 Current address: Network Control Engineering, Akamai Technologies Inc., 8 Cambridge Center, Cambridge, MA 02142, USA
3 Current address: Engineering, Google Inc., 5 Cambridge Center, Cambridge, MA 02142, USA
4 Genome Sequencing and Analysis Program, Broad Institute of MIT & Harvard, 7 Cambridge Center, Cambridge, MA 02142, USA
5 Current address: Genomic Technologies, Joint Genome Institute, Walnut Creek, CA 94598, USA
Genome Biology 2010, 11:R15 doi:10.1186/gb-2010-11-2-r15
Published: 5 February 2010Abstract
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.