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

Genome Biol. 2010;11(2):R15. doi: 10.1186/gb-2010-11-2-r15. Epub 2010 Feb 5.


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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Algorithms
  • Electronic Data Processing*
  • Gene Library*
  • High-Throughput Screening Assays*
  • Humans
  • Microspheres
  • Sequence Analysis, DNA / methods*