Publication Date:
2008-04-05
Description:
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.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Harris, Timothy D -- Buzby, Phillip R -- Babcock, Hazen -- Beer, Eric -- Bowers, Jayson -- Braslavsky, Ido -- Causey, Marie -- Colonell, Jennifer -- Dimeo, James -- Efcavitch, J William -- Giladi, Eldar -- Gill, Jaime -- Healy, John -- Jarosz, Mirna -- Lapen, Dan -- Moulton, Keith -- Quake, Stephen R -- Steinmann, Kathleen -- Thayer, Edward -- Tyurina, Anastasia -- Ward, Rebecca -- Weiss, Howard -- Xie, Zheng -- R01 HG004144-01/HG/NHGRI NIH HHS/ -- Howard Hughes Medical Institute/ -- New York, N.Y. -- Science. 2008 Apr 4;320(5872):106-9. doi: 10.1126/science.1150427.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Helicos BioSciences Corporation, One Kendall Square, Cambridge, MA 02139, USA. tharris@helicosbio.com〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/18388294" target="_blank"〉PubMed〈/a〉
Keywords:
Algorithms
;
Bacteriophage M13/*genetics
;
Computational Biology/methods
;
DNA Primers
;
DNA, Viral/chemistry/*genetics
;
*Genome, Viral
;
Mutation
;
Sequence Alignment
;
Sequence Analysis, DNA/*methods
;
Software
;
Templates, Genetic
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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