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

Autor:	Boskovic N; Department of Biosciences and Nutrition, Karolinska Institutet, 14183 Huddinge, Sweden; Department of Obstetrics and Gynecology, University of Helsinki, 00290 Helsinki, Finland. Electronic address: nina.boskovic@ki.se., Yazgeldi G; Folkhälsan Research Center, 00290 Helsinki, Finland; Stem Cells and Metabolism Research Program, University of Helsinki, 00290 Helsinki, Finland., Ezer S; Folkhälsan Research Center, 00290 Helsinki, Finland; Stem Cells and Metabolism Research Program, University of Helsinki, 00290 Helsinki, Finland., Tervaniemi MH; Folkhälsan Research Center, 00290 Helsinki, Finland; Stem Cells and Metabolism Research Program, University of Helsinki, 00290 Helsinki, Finland., Inzunza J; Department of Biosciences and Nutrition, Karolinska Institutet, 14183 Huddinge, Sweden., Deligiannis SP; Department of Obstetrics and Gynecology, University of Helsinki, 00290 Helsinki, Finland; Department of Obstetrics and Gynecology, Institute of Clinical Medicine, University of Tartu, 50406 Tartu, Estonia., Yaşar B; Department of Biosciences and Nutrition, Karolinska Institutet, 14183 Huddinge, Sweden; Department of Biotechnology, Institute of Molecular and Cell Biology, University of Tartu, 51010 Tartu, Estonia., Skoog T; Department of Biosciences and Nutrition, Karolinska Institutet, 14183 Huddinge, Sweden., Krjutškov K; Competence Centre of Health Technologies, 50411 Tartu, Estonia; Department of Obstetrics and Gynecology, Institute of Clinical Medicine, University of Tartu, 50406 Tartu, Estonia., Katayama S; Department of Biosciences and Nutrition, Karolinska Institutet, 14183 Huddinge, Sweden; Folkhälsan Research Center, 00290 Helsinki, Finland; Stem Cells and Metabolism Research Program, University of Helsinki, 00290 Helsinki, Finland. Electronic address: shintaro.katayama@folkhalsan.fi., Kere J; Department of Biosciences and Nutrition, Karolinska Institutet, 14183 Huddinge, Sweden; Folkhälsan Research Center, 00290 Helsinki, Finland; Stem Cells and Metabolism Research Program, University of Helsinki, 00290 Helsinki, Finland. Electronic address: juha.kere@ki.se.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2023 Sep 15; Vol. 4 (3), pp. 102357. Date of Electronic Publication: 2023 Jun 13.
DOI:	10.1016/j.xpro.2023.102357
Abstrakt:	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 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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