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Báo cáo y học: " A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học Wertheim cung cấp cho các bạn kiến thức về ngành y đề tài: A scalable, fully automated process for construction of sequence-ready barcoded libraries for 454. | Lennon et al. Genome Biology 2010 11 R15 http genomebiology.eom 2010 11 2 R15 w Genome Biology METHOD Open Access A sealable fully automated process for construction of sequenee-ready barcoded libraries for 454 Niall J Lennon 1 Robert E Lintner1 Scott Anderson1 Pablo Alvarez2 Andrew Barry1 William Brockman3 Riza Daza1 Rachel L Erlich1 Georgia Giannoukos4 Lisa Green1 Andrew Hollinger1 Cindi A Hoover5 David B Jaffe4 Frank Juhn1 Danielle McCarthy1 Danielle Perrin1 Karen Ponchner1 Taryn L Powers1 Kamran Rizzolo1 Dana Robbins1 Elizabeth Ryan1 Carsten Russ4 Todd Sparrow1 John Stalker1 Scott Steelman1 Michael Weiand1 Andrew Zimmer1 Matthew R Henn1 Chad Nusbaum4 and Robert Nicol 1 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. Background The emergence of next-generation sequencing technologies such as the Roche 454 Genome Sequencer the Illumina Genome Analyzer the Applied Biosystems SOLiD sequencer and others has provided the opportunity for both large genome centers and individual labs to generate DNA sequence data at an unprecedented scale 1 . However as sequence output continues to increase dramatically processes to generate sequence-ready libraries lag behind in scale. The minimum unit of sequence data for example lane or channel already exceeds the amount required for small projects such as viral or bacterial genomes and will continue to increase. As a result projects with large numbers of samples but small sequence per sample requirements become increasingly challenging to .