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Accurate whole-genome sequencing and haplotyping from 10 to 20 human cells

2012; Nature Portfolio; Volume: 487; Issue: 7406 Linguagem: Inglês

10.1038/nature11236

1476-4687

Brock A. Peters, Bahram G. Kermani, Andrew B. Sparks, Oleg Alferov, Peter Hong, Andrei Alexeev, Yuan Jiang, Fredrik A. Dahl, Yanqing Tang, Juergen Haas, Kimberly Robasky, Alexander Wait Zaranek, Je‐Hyuk Lee, Mad Price Ball, Joseph E. Peterson, Helena Perazich, George Yeung, Jia Liu, Linsu Chen, Michael Kennemer, Kaliprasad Pothuraju, Karel Konvicka, Mike Tsoupko-Sitnikov, Krishna Prasad Pant, Jessica Ebert, Geoffrey B. Nilsen, Jonathan Baccash, Aaron L. Halpern, George M. Church, Radoje Drmanac,

Cancer Genomics and Diagnostics

Recent advances in whole-genome sequencing have brought the vision of personal genomics and genomic medicine closer to reality. However, current methods lack clinical accuracy and the ability to describe the context (haplotypes) in which genome variants co-occur in a cost-effective manner. Here we describe a low-cost DNA sequencing and haplotyping process, long fragment read (LFR) technology, which is similar to sequencing long single DNA molecules without cloning or separation of metaphase chromosomes. In this study, ten LFR libraries were made using only ∼100 picograms of human DNA per sample. Up to 97% of the heterozygous single nucleotide variants were assembled into long haplotype contigs. Removal of false positive single nucleotide variants not phased by multiple LFR haplotypes resulted in a final genome error rate of 1 in 10 megabases. Cost-effective and accurate genome sequencing and haplotyping from 10-20 human cells, as demonstrated here, will enable comprehensive genetic studies and diverse clinical applications.