Single-Molecule DNA Sequencing of a Viral Genome

by: Timothy D Harris, Phillip R Buzby, Hazen Babcock, Eric Beer, Jayson Bowers, Ido Braslavsky, Marie Causey, Jennifer Colonell, James Dimeo, William J Efcavitch, Eldar Giladi, Jaime Gill, John Healy, Mirna Jarosz, Dan Lapen, Keith Moulton, Stephen R Quake, Kathleen Steinmann, Edward Thayer, Anastasia Tyurina, Rebecca Ward, Howard Weiss, Zheng Xie

Science, Vol. 320, No. 5872. (4 April 2008), pp. 106-109.

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摘要

The full promise of human genomics will be realized only when the genomes of thousands of individuals can be sequenced for comparative analysis. A reference sequence enables the use of short read length. We report an amplification-free method for determining the nucleotide sequence of more than 280,000 individual DNA molecules simultaneously. A DNA polymerase adds labeled nucleotides to surface-immobilized primer-template duplexes in stepwise fashion, and the asynchronous growth of individual DNA molecules was monitored by fluorescence imaging. Read lengths of >25 bases and equivalent phred software program quality scores approaching 30 were achieved. We used this method to sequence the M13 virus to an average depth of >150x and with 100% coverage; thus, we resequenced the M13 genome with high-sensitivity mutation detection. This demonstrates a strategy for high-throughput low-cost resequencing. 10.1126/science.1150427

@article{citeulike:2630042, abstract = {The full promise of human genomics will be realized only when the genomes of thousands of individuals can be sequenced for comparative analysis. A reference sequence enables the use of short read length. We report an amplification-free method for determining the nucleotide sequence of more than 280,000 individual DNA molecules simultaneously. A DNA polymerase adds labeled nucleotides to surface-immobilized primer-template duplexes in stepwise fashion, and the asynchronous growth of individual DNA molecules was monitored by fluorescence imaging. Read lengths of >25 bases and equivalent phred software program quality scores approaching 30 were achieved. We used this method to sequence the M13 virus to an average depth of >150x and with 100\% coverage; thus, we resequenced the M13 genome with high-sensitivity mutation detection. This demonstrates a strategy for high-throughput low-cost resequencing. 10.1126/science.1150427}, author = {Harris, Timothy D. and Buzby, Phillip R. and Babcock, Hazen and Beer, Eric and Bowers, Jayson and Braslavsky, Ido and Causey, Marie and Colonell, Jennifer and Dimeo, James and Efcavitch, William J. and Giladi, Eldar and Gill, Jaime and Healy, John and Jarosz, Mirna and Lapen, Dan and Moulton, Keith and Quake, Stephen R. and Steinmann, Kathleen and Thayer, Edward and Tyurina, Anastasia and Ward, Rebecca and Weiss, Howard and Xie, Zheng }, citeulike-article-id = {2630042}, doi = {http://dx.doi.org/10.1126/science.1150427}, journal = {Science}, month = {April}, number = {5872}, pages = {106--109}, posted-at = {2008-06-16 16:33:47}, priority = {2}, title = {Single-Molecule DNA Sequencing of a Viral Genome}, url = {http://dx.doi.org/10.1126/science.1150427}, volume = {320}, year = {2008} }

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TY - JOUR ID - citeulike:2630042 L3 - citeulike-article-id:2630042 N2 - The full promise of human genomics will be realized only when the genomes of thousands of individuals can be sequenced for comparative analysis. A reference sequence enables the use of short read length. We report an amplification-free method for determining the nucleotide sequence of more than 280,000 individual DNA molecules simultaneously. A DNA polymerase adds labeled nucleotides to surface-immobilized primer-template duplexes in stepwise fashion, and the asynchronous growth of individual DNA molecules was monitored by fluorescence imaging. Read lengths of >25 bases and equivalent phred software program quality scores approaching 30 were achieved. We used this method to sequence the M13 virus to an average depth of >150x and with 100% coverage; thus, we resequenced the M13 genome with high-sensitivity mutation detection. This demonstrates a strategy for high-throughput low-cost resequencing. 10.1126/science.1150427 IS - 5872 JF - Science EP - 109 TI - Single-Molecule DNA Sequencing of a Viral Genome VL - 320 SP - 106 AU - Harris, Timothy AU - Buzby, Phillip AU - Babcock, Hazen AU - Beer, Eric AU - Bowers, Jayson AU - Braslavsky, Ido AU - Causey, Marie AU - Colonell, Jennifer AU - Dimeo, James AU - Efcavitch, William AU - Giladi, Eldar AU - Gill, Jaime AU - Healy, John AU - Jarosz, Mirna AU - Lapen, Dan AU - Moulton, Keith AU - Quake, Stephen AU - Steinmann, Kathleen AU - Thayer, Edward AU - Tyurina, Anastasia AU - Ward, Rebecca AU - Weiss, Howard AU - Xie, Zheng PY - 2008/04/04/ UR - http://dx.doi.org/10.1126/science.1150427 ER -

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