Single-molecule sequencing of an individual human genome Export

@article{citeulike:5414362, abstract = {Recent advances in high-throughput DNA sequencing technologies have enabled order-of-magnitude improvements in both cost and throughput. Here we report the use of single-molecule methods to sequence an individual human genome. We aligned billions of 24- to 70-bp reads (32 bp average) to 90\% of the National Center for Biotechnology Information (NCBI) reference genome, with 28 average coverage. Our results were obtained on one sequencing instrument by a single operator with four data collection runs. Single-molecule sequencing enabled analysis of human genomic information without the need for cloning, amplification or ligation. We determined 2.8 million single nucleotide polymorphisms (SNPs) with a false-positive rate of less than 1\% as validated by Sanger sequencing and 99.8\% concordance with SNP genotyping arrays. We identified 752 regions of copy number variation by analyzing coverage depth alone and validated 27 of these using digital PCR. This milestone should allow widespread application of genome sequencing to many aspects of genetics and human health, including personal genomics.}, author = {Pushkarev, Dmitry and Neff, Norma F. and Quake, Stephen R.}, citeulike-article-id = {5414362}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nbt.1561}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nbt.1561}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/19668243}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=19668243}, comment = {28-fold coverage}, day = {10}, doi = {10.1038/nbt.1561}, issn = {1087-0156}, journal = {Nature Biotechnology}, keywords = {3rd-generation, genome, helicos, individual-human-genome, single-molecule}, month = {August}, number = {9}, pages = {847--850}, posted-at = {2009-10-28 23:04:32}, priority = {2}, publisher = {Nature Publishing Group}, title = {Single-molecule sequencing of an individual human genome}, url = {http://dx.doi.org/10.1038/nbt.1561}, volume = {27}, year = {2009} }

TY - JOUR ID - citeulike:5414362 L3 - citeulike-article-id:5414362 N2 - Recent advances in high-throughput DNA sequencing technologies have enabled order-of-magnitude improvements in both cost and throughput. Here we report the use of single-molecule methods to sequence an individual human genome. We aligned billions of 24- to 70-bp reads (32 bp average) to 90% of the National Center for Biotechnology Information (NCBI) reference genome, with 28 average coverage. Our results were obtained on one sequencing instrument by a single operator with four data collection runs. Single-molecule sequencing enabled analysis of human genomic information without the need for cloning, amplification or ligation. We determined 2.8 million single nucleotide polymorphisms (SNPs) with a false-positive rate of less than 1% as validated by Sanger sequencing and 99.8% concordance with SNP genotyping arrays. We identified 752 regions of copy number variation by analyzing coverage depth alone and validated 27 of these using digital PCR. This milestone should allow widespread application of genome sequencing to many aspects of genetics and human health, including personal genomics. IS - 9 JF - Nature Biotechnology SN - 1087-0156 EP - 850 TI - Single-molecule sequencing of an individual human genome PB - Nature Publishing Group VL - 27 SP - 847 KW - 3rd-generation KW - genome KW - helicos KW - individual-human-genome KW - single-molecule N1 - 28-fold coverage AU - Pushkarev, Dmitry AU - Neff, Norma AU - Quake, Stephen PY - 2009/08/10/ UR - http://dx.doi.org/10.1038/nbt.1561 ER -