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http://223.31.159.10:8080/jspui/handle/123456789/1154
Title: | Single cell type specific RNA isolation and gene expression analysis in rice using laser capture microdissection (LCM)-based method |
Authors: | Gautam, Vibhav Chatterjee, Sourav Sarkar, Ananda K. |
Keywords: | LCM Laser microdissection RNA isolation Gene expression Mesophyll cells Bundle sheath cells Rice |
Issue Date: | 2021 |
Publisher: | Springer Nature Publishing AG |
Citation: | Methods in Molecular Biology, 2238: 275-283 |
Abstract: | The success of single cell type-specific gene expression or functional study largely depends on the efficient isolation of high-quality RNA from them. Laser capture microdissection (LCM) is an efficient technique that allows accessing and dissecting out a specific individual cell or cell type from a microscopic heterogeneous tissue in a minimally disruptive way. Here, we describe an efficient and inexpensive LCM-based method for the extraction of RNAs with high yield and integrity from laser-microdissected mesophyll and bundle sheath cells of rice leaf. The integrity of isolated RNA is assessed with bioanalyzer analysis, and the presence of mRNA of a specific gene is validated through RT-PCR. This RNA could further be used for uncovering single cell type-specific gene expression signature using next-generation transcriptome sequence or through regular RT-PCR. |
Description: | Accepted date: 21 January 2021 |
URI: | https://link.springer.com/protocol/10.1007%2F978-1-0716-1068-8_18 http://223.31.159.10:8080/jspui/handle/123456789/1154 |
ISBN: | 978-1-0716-1067-1 |
Appears in Collections: | Institutional Publications |
Files in This Item:
File | Description | Size | Format | |
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Sarkar AK_2021_1.pdf Restricted Access | 380.31 kB | Adobe PDF | View/Open Request a copy |
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