Protocol for high-quality single-cell RNA-seq from tissue sections with DRaqL.

Autor: Ikeda H; Department of Embryology, School of Medicine, Nara Medical University, Kashihara, Nara 634-8521, Japan., Miyao S; Department of Embryology, School of Medicine, Nara Medical University, Kashihara, Nara 634-8521, Japan., Yamada N; Department of Embryology, School of Medicine, Nara Medical University, Kashihara, Nara 634-8521, Japan., Sugimoto S; Department of Obstetrics and Gynecology, School of Medicine, Nara Medical University, Kashihara, Nara 634-8521, Japan., Kimura F; Department of Obstetrics and Gynecology, School of Medicine, Nara Medical University, Kashihara, Nara 634-8521, Japan., Kurimoto K; Department of Embryology, School of Medicine, Nara Medical University, Kashihara, Nara 634-8521, Japan. Electronic address: kurimoto@naramed-u.ac.jp.
Jazyk: angličtina
Zdroj: STAR protocols [STAR Protoc] 2024 Jun 21; Vol. 5 (2), pp. 103050. Date of Electronic Publication: 2024 May 02.
DOI: 10.1016/j.xpro.2024.103050
Abstrakt: Single-cell RNA sequencing (scRNA-seq) combined with laser capture microdissection (LCM) offers a versatile framework for comprehensive transcriptomics from tissue sections. Here, we present a detailed protocol for DRaqL (direct RNA recovery and quenching for LCM) in combination with Smart-seq2 (DRaqL-Smart-seq2), which enables high-quality RNA sequencing for single cells obtained from alcohol-fixed murine ovarian sections. Additionally, we provide an optional procedure for scRNA-seq from formalin-fixed sections (DRaqL-Protease-Smart-seq2). We outline key steps for cell lysis, cDNA amplification, and sequencing library preparation. For complete details on the use and execution of this protocol, please refer to Ikeda et al. 1 .
Competing Interests: Declaration of interests H.K., S.M., and K.K. have a patent pending in Japan on DRaqL (application number 2022-194046).
(Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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