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PMID: 19668243 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Single-molecule sequencing of an individual human genome.

Nature biotechnology ·Vol. 27 ·No. 9 ·2009-09-00 ·Pages 847-50

Pushkarev D, Neff NF, Quake SR

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 approximately 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 approximately 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.

MeSH Terms
Computer Simulation Genome, Human Genomics/methods Humans Polymorphism, Single Nucleotide Reproducibility of Results Sequence Analysis, DNA/methods
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Pushkarev Dmitry
Department of Bioengineering, Stanford University and Howard Hughes Medical Institute, Stanford, California, USA.
Neff Norma F
Quake Stephen R
References (21)
21 references, click to expand
  1. Aging and genome maintenance.
    Ann N Y Acad Sci. 2005 Dec;1055:35-47 PMID: 16387716
  2. Filter-based hybridization capture of subgenomes enables resequencing and copy-number detection.
    Nat Methods. 2009 Jul;6(7):507-10 PMID: 19543287
  3. Studying copy number variations using a nanofluidic platform.
    Nucleic Acids Res. 2008 Oct;36(18):e116 PMID: 18710881
  4. The first Korean genome sequence and analysis: full genome sequencing for a socio-ethnic group.
    Genome Res. 2009 Sep;19(9):1622-9 PMID: 19470904
  5. High-throughput multiplex sequencing to discover copy number variants in Drosophila.
    Genetics. 2009 Aug;182(4):935-41 PMID: 19528327
  6. The diploid genome sequence of an Asian individual.
    Nature. 2008 Nov 6;456(7218):60-5 PMID: 18987735
  7. A highly annotated whole-genome sequence of a Korean individual.
    Nature. 2009 Aug 20;460(7258):1011-5 PMID: 19587683
  8. The sequence of the human genome.
    Science. 2001 Feb 16;291(5507):1304-51 PMID: 11181995
  9. Mapping short DNA sequencing reads and calling variants using mapping quality scores.
    Genome Res. 2008 Nov;18(11):1851-8 PMID: 18714091
  10. Detection of large-scale variation in the human genome.
    Nat Genet. 2004 Sep;36(9):949-51 PMID: 15286789
  11. Real-time DNA sequencing from single polymerase molecules.
    Science. 2009 Jan 2;323(5910):133-8 PMID: 19023044
  12. CNV-seq, a new method to detect copy number variation using high-throughput sequencing.
    BMC Bioinformatics. 2009 Mar 06;10:80 PMID: 19267900
  13. Sequence information can be obtained from single DNA molecules.
    Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):3960-4 PMID: 12651960
  14. Single-molecule, motion-based DNA sequencing using RNA polymerase.
    Science. 2006 Aug 11;313(5788):801 PMID: 16902131
  15. The complete genome of an individual by massively parallel DNA sequencing.
    Nature. 2008 Apr 17;452(7189):872-6 PMID: 18421352
  16. SHRiMP: accurate mapping of short color-space reads.
    PLoS Comput Biol. 2009 May;5(5):e1000386 PMID: 19461883
  17. The diploid genome sequence of an individual human.
    PLoS Biol. 2007 Sep 4;5(10):e254 PMID: 17803354
  18. Accurate whole human genome sequencing using reversible terminator chemistry.
    Nature. 2008 Nov 6;456(7218):53-9 PMID: 18987734
  19. Single-molecule DNA sequencing of a viral genome.
    Science. 2008 Apr 4;320(5872):106-9 PMID: 18388294
  20. Initial sequencing and analysis of the human genome.
    Nature. 2001 Feb 15;409(6822):860-921 PMID: 11237011
  21. DNA sequencing of a cytogenetically normal acute myeloid leukaemia genome.
    Nature. 2008 Nov 6;456(7218):66-72 PMID: 18987736
Article Info
Journal
Nature biotechnology
Abbr.
Nat Biotechnol
ISSN
1546-1696
Published
2009-09-00
Epub
2009-00-10
Pages
847-50
Language
English
Region
United States
NLM ID
9604648
PMCID
PMC4117198
Subset
IM
Grants
NIH HHS · DP1 OD000251 · United States
Corrections
CommentIn
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