semi-automated library preparation for high-throughput dna sequencing platforms半自动图书馆准备高通量dna测序平台.pdfVIP

Hindawi Publishing Corporation Journal of Biomedicine and Biotechnology Volume 2010, Article ID 617469, 8 pages doi:10.1155/2010/617469 Methodology Report Semi-Automated Library Preparation for High-Throughput DNA Sequencing Platforms Eveline Farias-Hesson,1 Jonathan Erikson,1 Alexander Atkins,1 Peidong Shen,2 Ronald W. Davis,2 Curt Scharfe,2 and Nader Pourmand1 1 Department of Biomolecular Engineering, University of California, 1156 High Street, Santa Cruz, CA 95064, USA 2 Stanford GenomeTechnology Center, Stanford University, 855 S. California Avenue, Palo Alto, CA 94304, USA Correspondence should be addressed to Nader Pourmand, pourmand@ Received 26 January 2010; Accepted 5 April 2010 Academic Editor: Lori Snyder Copyright © 2010 Eveline Farias-Hesson et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Next-generation sequencing platforms are powerful technologies, providing gigabases of genetic information in a single run. An important prerequisite for high-throughput DNA sequencing is the development of robust and cost-effective preprocessing protocols for DNA sample library construction. Here we report the development of a semi-automated sample preparation protocol to produce adaptor-ligated fragment libraries. Using a liquid-handling robot in conjunction with Carboxy Terminated Magnetic Beads, we labeled each library sample using a unique 6 bp DNA barcode, which allowed multiplex sample processing and sequencing of 32 libraries in a single run using Applied Biosystems’ SOLiD sequencer. We applied our semi-automated pipeline to targeted medical resequencing