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Improved method of RNA isolation from laser capture microdissection (LCM)-derived plant tissues

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Title Improved method of RNA isolation from laser capture microdissection (LCM)-derived plant tissues
 
Creator Gautam, Vibhav
Singh, Archita
Singh, Sharmila
Verma, Swati
Sarkar, Ananda K.
 
Subject LCM
Laser microdissection
RNA isolation
Gene expression
miRNA
Root meristem
 
Description Accepted date: 04 April 2019
Laser capture microdissection (LCM) is a tool to isolate desired and/or less accessible cells or tissues from a heterogeneous population. In the current method, we describe an efficient and cost-effective method to obtain both high-quality mRNA and miRNAs in sufficient quantity from LCM-derived plant tissues. The quality of the isolated RNA can be assessed using Bioanalyzer. Using modified stem-loop RT-PCR, we confirmed the presence of 21–24 nucleotide (nt) long mature miRNAs. This modified LCM-based method has been found to be suitable for the tissue-specific expression analysis of both genes and small RNAs (miRNAs).
V.G., A.S., and S.S. thank the Council for Scientific and Industrial Research (CSIR), India, and the National Institute of Plant Genome Research (NIPGR), New Delhi, India, for funding and internal grants. V.G. and A.S. also acknowledge the Department of Biotechnology (DBT), India, for fellowship. S.V. thanks the Department of Science and Technology-Science and Engineering Research Board (DST-SERB), India, for National Post Doctoral Fellowship (N-PDF). A.K.S. thanks NIPGR and DBT (Project Grant No. BT/PR12766/BPA/118/63/2015), New Delhi, India, for fellowship and grants. The authors declare no conflict of interests.
 
Date 2019-04-11T10:22:19Z
2019-04-11T10:22:19Z
2019
 
Type Book chapter
 
Identifier Methods in Molecular Biology, 1933: 89-98
978-1-4939-9045-0
http://223.31.159.10:8080/jspui/handle/123456789/940
https://link.springer.com/protocol/10.1007%2F978-1-4939-9045-0_5
10.1007/978-1-4939-9045-0_5.
 
Language en_US
 
Format application/pdf
 
Publisher Springer Nature