High-Throughput Single-Cell Sequencing with Linear Amplification.
Mol Cell
; 76(4): 676-690.e10, 2019 11 21.
Article
em En
| MEDLINE
| ID: mdl-31495564
ABSTRACT
Conventional methods for single-cell genome sequencing are limited with respect to uniformity and throughput. Here, we describe sci-L3, a single-cell sequencing method that combines combinatorial indexing (sci-) and linear (L) amplification. The sci-L3 method adopts a 3-level (3) indexing scheme that minimizes amplification biases while enabling exponential gains in throughput. We demonstrate the generalizability of sci-L3 with proof-of-concept demonstrations of single-cell whole-genome sequencing (sci-L3-WGS), targeted sequencing (sci-L3-target-seq), and a co-assay of the genome and transcriptome (sci-L3-RNA/DNA). We apply sci-L3-WGS to profile the genomes of >10,000 sperm and sperm precursors from F1 hybrid mice, mapping 86,786 crossovers and characterizing rare chromosome mis-segregation events in meiosis, including instances of whole-genome equational chromosome segregation. We anticipate that sci-L3 assays can be applied to fully characterize recombination landscapes, to couple CRISPR perturbations and measurements of genome stability, and to other goals requiring high-throughput, high-coverage single-cell sequencing.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Análise de Sequência de RNA
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Análise de Sequência de DNA
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Perfilação da Expressão Gênica
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Técnicas de Amplificação de Ácido Nucleico
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Análise de Célula Única
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Sequenciamento de Nucleotídeos em Larga Escala
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Sequenciamento Completo do Genoma
Limite:
Animals
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article