Optimized single-cell RNA sequencing protocol to study early genome activation in mammalian preimplantation development

被引：5

作者：

Boskovic, Nina ^{[1
,2
]}

Yazgeldi, Gamze ^{[3
,4
]}

Ezer, Sini ^{[3
,4
]}

Tervaniemi, Mari H. ^{[3
,4
]}

Inzunza, Jose ^{[1
]}

Deligiannis, Spyridon Panagiotis ^{[2
]}

Yasar, Baris ^{[1
,7
]}

Skoog, Tiina ^{[1
]}

Krjutskov, Kaarel ^{[5
,6
]}

Katayama, Shintaro ^{[1
,3
,4
]}

Kere, Juha ^{[1
,3
,4
]}

机构：

[1] Karolinska Inst, Dept Biosci & Nutr, S-14183 Huddinge, Sweden

[2] Univ Helsinki, Dept Obstet & Gynecol, Helsinki 00290, Finland

[3] Folkhalsan Res Ctr, Helsinki 00290, Finland

[4] Univ Helsinki, Stem Cells & Metab Res Program, FI-00290 Helsinki, Finland

[5] Competence Ctr Hlth Technol, EE-50410 Tartu, Estonia

[6] Univ Tartu, Inst Clin Med, Dept Obstet & Gynecol, EE-50406 Tartu, Estonia

[7] Univ Tartu, Inst Mol & Cell Biol, Dept Biotechnol, EE-51010 Tartu, Estonia

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 03期

基金：

瑞典研究理事会;

关键词：

Bioinformatics; Developmental biology; Gene Expression; Genomics; Molecular Biology; RNA-seq; Sequence analysis; Sequencing; Single Cell;

D O I：

10.1016/j.xpro.2023.102357

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Here, we present a modification of single-cell tagged reverse transcription protocol to study gene expression on a single-cell level or with limited RNA input. We describe different enzymes for reverse transcription and cDNA amplification, modified lysis buffer, and additional clean-up steps before cDNA amplification. We also detail an optimized single-cell RNA sequencing method for handpicked single cells, or tens to hundreds of cells, as input material to study mammalian preimplantation development. For complete details on the use and execution of this protocol, please refer to Ezer et al.1

引用

页数：26

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