Genome-wide copy number analysis of single cells

被引：260

作者：

Baslan, Timour ^{[1
,2
]}

Kendall, Jude ^{[1
]}

Rodgers, Linda ^{[1
]}

Cox, Hilary ^{[1
]}

Riggs, Mike ^{[1
]}

Stepansky, Asya ^{[1
]}

Troge, Jennifer ^{[1
]}

Ravi, Kandasamy ^{[1
]}

Esposito, Diane ^{[1
]}

Lakshmi, B. ^{[3
]}

Wigler, Michael ^{[1
]}

Navin, Nicholas ^{[4
,5
]}

Hicks, James ^{[1
]}

机构：

[1] Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA

[2] SUNY Stony Brook, Mol & Cellular Biol Program, Stony Brook, NY 11794 USA

[3] Ontario Inst Canc Res, Toronto, ON, Canada

[4] Univ Texas MD Anderson Canc Ctr, Dept Genet, Houston, TX 77030 USA

[5] Univ Texas MD Anderson Canc Ctr, Dept Bioinformat & Computat Biol, Houston, TX 77030 USA

来源：

NATURE PROTOCOLS | 2012年 / 7卷 / 06期

关键词：

ARRAY CGH; EVOLUTION; DNA; METASTASIS; RESOLUTION;

D O I：

10.1038/nprot.2012.039

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Copy number variation (CNV) is increasingly recognized as an important contributor to phenotypic variation in health and disease. Most methods for determining CNV rely on admixtures of cells in which information regarding genetic heterogeneity is lost. Here we present a protocol that allows for the genome-wide copy number analysis of single nuclei isolated from mixed populations of cells. Single-nucleus sequencing (SNS), combines flow sorting of single nuclei on the basis of DNA content and whole-genome amplification (WGA); this is followed by next-generation sequencing to quantize genomic intervals in a genome-wide manner. Multiplexing of single cells is discussed. In addition, we outline informatic approaches that correct for biases inherent in the WGA procedure and allow for accurate determination of copy number profiles. All together, the protocol takes similar to 3 d from flow cytometry to sequence-ready DNA libraries.

引用

页码：1024 / 1041

页数：18

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