Genome sequencing in microfabricated high-density picolitre reactors Export

@article{margulies05, abstract = {The proliferation of large-scale DNA-sequencing projects in recent years has driven a search for alternative methods to reduce time and cost. Here we describe a scalable, highly parallel sequencing system with raw throughput significantly greater than that of state-of-the-art capillary electrophoresis instruments. The apparatus uses a novel fibre-optic slide of individual wells and is able to sequence 25 million bases, at 99\% or better accuracy, in one four-hour run. To achieve an approximately 100-fold increase in throughput over current Sanger sequencing technology, we have developed an emulsion method for DNA amplification and an instrument for sequencing by synthesis using a pyrosequencing protocol optimized for solid support and picolitre-scale volumes. Here we show the utility, throughput, accuracy and robustness of this system by shotgun sequencing and de novo assembly of the Mycoplasma genitalium genome with 96\% coverage at 99.96\% accuracy in one run of the machine.}, author = {Margulies, Marcel and Egholm, Michael and Altman, William E. and Attiya, Said and Bader, Joel S. and Bemben, Lisa A. and Berka, Jan and Braverman, Michael S. and Chen, Yi J. and Chen, Zhoutao and Dewell, Scott B. and Du, Lei and Fierro, Joseph M. and Gomes, Xavier V. and Godwin, Brian C. and He, Wen and Helgesen, Scott and Ho, Chun H. and Irzyk, Gerard P. and Jando, Szilveszter C. and Alenquer, Maria L. I. and Jarvie, Thomas P. and Jirage, Kshama B. and Kim, Jong B. and Knight, James R. and Lanza, Janna R. and Leamon, John H. and Lefkowitz, Steven M. and Lei, Ming and Li, Jing and Lohman, Kenton L. and Lu, Hong and Makhijani, Vinod B. and Mcdade, Keith E. and Mckenna, Michael P. and Myers, Eugene W. and Nickerson, Elizabeth and Nobile, John R. and Plant, Ramona and Puc, Bernard P. and Ronan, Michael T. and Roth, George T. and Sarkis, Gary J. and Simons, Jan F. and Simpson, John W. and Srinivasan, Maithreyan and Tartaro, Karrie R. and Tomasz, Alexander and Vogt, Kari A. and Volkmer, Greg A. and Wang, Shally H. and Wang, Yong and Weiner, Michael P. and Yu, Pengguang and Begley1, Richard F. and Rothberg, Jonathan M.}, citeulike-article-id = {1570525}, doi = {doi:10.1038/nature03959}, journal = {Nature}, month = {September}, number = {7057}, pages = {376--380}, posted-at = {2007-08-16 21:27:03}, priority = {0}, title = {Genome sequencing in microfabricated high-density picolitre reactors}, volume = {437}, year = {2005} }

TY - JOUR ID - margulies05 L3 - citeulike-article-id:1570525 N2 - The proliferation of large-scale DNA-sequencing projects in recent years has driven a search for alternative methods to reduce time and cost. Here we describe a scalable, highly parallel sequencing system with raw throughput significantly greater than that of state-of-the-art capillary electrophoresis instruments. The apparatus uses a novel fibre-optic slide of individual wells and is able to sequence 25 million bases, at 99\% or better accuracy, in one four-hour run. To achieve an approximately 100-fold increase in throughput over current Sanger sequencing technology, we have developed an emulsion method for DNA amplification and an instrument for sequencing by synthesis using a pyrosequencing protocol optimized for solid support and picolitre-scale volumes. Here we show the utility, throughput, accuracy and robustness of this system by shotgun sequencing and de novo assembly of the Mycoplasma genitalium genome with 96\% coverage at 99.96\% accuracy in one run of the machine. IS - 7057 JF - Nature EP - 380 TI - Genome sequencing in microfabricated high-density picolitre reactors VL - 437 SP - 376 AU - Margulies, Marcel AU - Egholm, Michael AU - Altman, William AU - Attiya, Said AU - Bader, Joel AU - Bemben, Lisa AU - Berka, Jan AU - Braverman, Michael AU - Chen, Yi AU - Chen, Zhoutao AU - Dewell, Scott AU - Du, Lei AU - Fierro, Joseph AU - Gomes, Xavier AU - Godwin, Brian AU - He, Wen AU - Helgesen, Scott AU - Ho, Chun AU - Irzyk, Gerard AU - Jando, Szilveszter AU - Alenquer, Maria AU - Jarvie, Thomas AU - Jirage, Kshama AU - Kim, Jong AU - Knight, James AU - Lanza, Janna AU - Leamon, John AU - Lefkowitz, Steven AU - Lei, Ming AU - Li, Jing AU - Lohman, Kenton AU - Lu, Hong AU - Makhijani, Vinod AU - Mcdade, Keith AU - Mckenna, Michael AU - Myers, Eugene AU - Nickerson, Elizabeth AU - Nobile, John AU - Plant, Ramona AU - Puc, Bernard AU - Ronan, Michael AU - Roth, George AU - Sarkis, Gary AU - Simons, Jan AU - Simpson, John AU - Srinivasan, Maithreyan AU - Tartaro, Karrie AU - Tomasz, Alexander AU - Vogt, Kari AU - Volkmer, Greg AU - Wang, Shally AU - Wang, Yong AU - Weiner, Michael AU - Yu, Pengguang AU - Begley1, Richard AU - Rothberg, Jonathan PY - 2005/September// ER -