Single-molecule sequencing of an individual human genome

被引:299
作者
Pushkarev, Dmitry [1 ,2 ]
Neff, Norma F. [1 ,2 ]
Quake, Stephen R. [1 ,2 ]
机构
[1] Stanford Univ, Dept Bioengn, Stanford, CA 94305 USA
[2] Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
Copy number variations - False positive rates - Genome sequencing - Genomic information - National center for biotechnology informations - Sanger sequencing - Single nucleotide polymorphisms - Single-molecule methods;
D O I
10.1038/nbt.1561
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 similar to 90% of the National Center for Biotechnology Information (NCBI) reference genome, with 28x 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 similar to 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.
引用
收藏
页码:847 / U101
页数:6
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