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April 13, 2017

[Report] Single-cell whole-genome analyses by Linear Amplification via Transposon Insertion (LIANTI)

ScienceNOW - Fetched: April 13th, 2017, 3:00pm UTC
Single-cell genomics is important for biology and medicine. However, current whole-genome amplification (WGA) methods are limited by low accuracy of copy-number variation (CNV) detection and low amplification fidelity. Here we report an improved single-cell WGA method, Linear Amplification via Transposon Insertion (LIANTI), which outperforms existing methods, enabling micro-CNV detection with kilobase resolution. This allowed direct observation of stochastic firing of DNA replication origins, which differs from cell to cell. We also show that the predominant cytosine-to-thymine mutations observed in single-cell genomics often arise from the artifact of cytosine deamination upon cell lysis. However, identifying single-nucleotide variations (SNVs) can be accomplished by sequencing kindred cells. We determined the spectrum of SNVs in a single human cell after ultraviolet radiation, revealing their nonrandom genome-wide distribution. Authors: Chongyi Chen, Dong Xing, Longzhi Tan, Heng Li, Guangyu Zhou, Lei Huang, X. Sunney Xie

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