Single-Cell Tagged Reverse Transcription (STRT-Seq)
Single-Cell Tagged Reverse Transcription (STRT-Seq)
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DOI：
10.1007/978-1-4939-9240-9_9
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发表时间：
2019-01-01
期刊：
影响因子：
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通讯作者：
Natarajan, Kedar Nath
中科院分区：
文献类型：
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作者：
Natarajan, Kedar Nath
单细胞RNA测序（scRNA-seq）已成为一种成熟的方法，用于分析来自不同细胞类型、组织、物种和生物体的单个细胞的整个转录组。单细胞标记的逆转录测序（STRT-seq）是早期的单细胞方法之一，其利用转录物的50标签计数。在Microfluidics Fluidigm C1平台（STRT-C1）上进行的STRT-seq是一种灵活的scRNA-seq方法，可以在细胞水平上对转录物进行准确，灵敏和重要的分子计数。在本文中，我描述了STRT-C1方法以及在C1微流控芯片上捕获96个细胞、cDNA合成和制备用于Illumina短读测序的单细胞文库所涉及的步骤。
Single-cell RNA sequencing (scRNA-seq) has become an established approach to profile entire transcriptomes of individual cells from different cell types, tissues, species, and organisms. Single-cell tagged reverse transcription sequencing (STRT-seq) is one of the early single-cell methods which utilize 5 0 tag counting of transcripts. STRT-seq performed on microfluidics Fluidigm C1 platform (STRT-C1) is a flexible scRNA-seq approach that allows for accurate, sensitive and importantly molecular counting of transcripts at singlecell level. Herein, I describe the STRT-C1 method and the steps involved in capturing 96 cells across C1 microfluidics chip, cDNA synthesis, and preparing single-cell libraries for Illumina short-read sequencing.