Improved laser capture microdissection (LCM)‑based method for isolation of RNA, including miRNA and expression analysis in woody apple bud meristem

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dc.contributor.authorVerma, Swati
dc.contributor.authorGautam, Vibhav
dc.contributor.authorSarkar, Ananda K.
dc.date.accessioned2019-04-18T10:52:42Z
dc.date.available2019-04-18T10:52:42Z
dc.date.issued2019
dc.descriptionAccepted date: 4 March 2019en_US
dc.description.abstractIsolation of high-quality RNA, including miRNA, from microscopic woody apple bud meristem using laser capture microdissection-based method. It is often challenging to study the expression of microRNAs (miRNAs) or genes in less accessible inner tissues of tree species rich in polyphenols or polysaccharides. Here, we report a laser capture microdissection (LCM)-based method for efficient and cost-effective isolation and expression analysis of miRNAs and genes in the meristem tissue of woody apple bud. The tissue fixation, processing, infiltration, and sectioning steps were optimized for LCM-based excision and subsequent RNA isolation. Further, we have confirmed that RNA isolated from LCM-derived apple bud meristem contained miRNAs and was of good quantity and quality, sufficient for downstream expression analysis.en_US
dc.description.sponsorshipSV acknowledges Department of Science and Technology-Science and Engineering Research Board (DST-SERB), Govt. of India, for National-Postdoctoral Fellowship (N-PDF fle no. PDF/2016/002423). AKS and VG acknowledge funding from Department of Biotechnology (DBT), Govt. of India (research grant no. BT/ PR12766/BPA/118/63/2015). We acknowledge LCM facility, other central instrument facility and internal grant of NIPGR. We thank Dr. Mohar Singh, ICAR-NBPGR, Regional Station Shimla, India for giving us access to collect apple bud samples. We thank the lab members (Shalini and Archita) for critical reading of the manuscript and valuable comments.en_US
dc.identifier.citationPlanta, 249(6): 2015-2020en_US
dc.identifier.doihttps://doi.org/10.1007/s00425-019-03127-0en_US
dc.identifier.issn1432-2048
dc.identifier.officialurlhttps://link.springer.com/article/10.1007%2Fs00425-019-03127-0en_US
dc.identifier.urihttps://ndkr-library.nipgr.ac.in/handle/123456789/944
dc.language.isoen_USen_US
dc.publisherSpringer Natureen_US
dc.subjectBud meristemen_US
dc.subjectGene expressionen_US
dc.subjectLaser capture microdissectionen_US
dc.subjectMalusen_US
dc.subjectMicroRNAsen_US
dc.subjectRNA isolationen_US
dc.subjectSmall RNAen_US
dc.subjectStem-loop RT-PCRen_US
dc.subjectTreeen_US
dc.titleImproved laser capture microdissection (LCM)‑based method for isolation of RNA, including miRNA and expression analysis in woody apple bud meristemen_US
dc.typeArticleen_US

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