使用Drop-Seq进行单细胞RNA测序。

Single-Cell RNA Sequencing with Drop-Seq.

作者信息

Bageritz Josephine, Raddi Gianmarco

机构信息

Division Signaling and Functional Genomics, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Wellcome Sanger Institute, University of Cambridge, Hinxton, UK.

出版信息

Methods Mol Biol. 2019;1979:73-85. doi: 10.1007/978-1-4939-9240-9_6.

DOI:10.1007/978-1-4939-9240-9_6

PMID:31028633

Abstract

Drop-Seq is a low-cost, high-throughput platform to profile thousands of cells by encapsualting them into individual droplets. Uniquely barcoded mRNA capture microparticles and cells are coconfined through a microfluidic device within the droplets where they undergo cell lysis and RNA hybridiztion. After breaking the droplets and pooling the hybridized particles, reverse transcription, PCR, and sequencing in single reactions allow to generate data from thousands of single-cell transcriptomes while maintaining information on the cellular origin of each transcript.

摘要

Drop-Seq是一个低成本、高通量的平台，通过将数千个细胞封装到单个液滴中来对其进行分析。带有独特条形码的mRNA捕获微粒和细胞通过微流控装置共同限制在液滴内，在液滴中它们会发生细胞裂解和RNA杂交。打破液滴并合并杂交后的微粒后，在单一反应中进行逆转录、PCR和测序，能够从数千个单细胞转录组中生成数据，同时保留每个转录本的细胞来源信息。