{"title":"Laser Microdissection–Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA","authors":"Divya Chandran, Michael J. Scanlon, Kazuhiro Ohtsu, Marja C.P. Timmermans, Patrick S. Schnable, Mary C. Wildermuth","doi":"10.1002/0471142727.mb25a03s112","DOIUrl":null,"url":null,"abstract":"<p>Laser microdissection of cells allows for isolation of specific cells of interest for downstream analyses including transcriptional profiling. Plant cells present unique challenges for laser microdissection due to their cellulosic cell walls and large vacuoles. Here we present protocols for plant tissue preparation, laser microdissection of select plant cells, and linear amplification of RNA from dissected cells. Linear amplification of RNA from dissected cells allows sufficient RNA for subsequent quantitative analysis by RT-PCR, microarray, or RNA sequencing. © 2015 by John Wiley & Sons, Inc.</p>","PeriodicalId":10734,"journal":{"name":"Current Protocols in Molecular Biology","volume":"112 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142727.mb25a03s112","citationCount":"22","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Molecular Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/0471142727.mb25a03s112","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 22
Abstract
Laser microdissection of cells allows for isolation of specific cells of interest for downstream analyses including transcriptional profiling. Plant cells present unique challenges for laser microdissection due to their cellulosic cell walls and large vacuoles. Here we present protocols for plant tissue preparation, laser microdissection of select plant cells, and linear amplification of RNA from dissected cells. Linear amplification of RNA from dissected cells allows sufficient RNA for subsequent quantitative analysis by RT-PCR, microarray, or RNA sequencing. © 2015 by John Wiley & Sons, Inc.
激光微解剖介导的植物RNA分离及体外转录扩增
激光显微解剖细胞允许分离特定细胞的下游分析,包括转录谱。植物细胞由于其纤维素细胞壁和大液泡,对激光显微解剖提出了独特的挑战。在这里,我们提出了植物组织制备,选择植物细胞的激光显微解剖,以及从解剖细胞中线性扩增RNA的方案。从解剖细胞的RNA线性扩增允许足够的RNA进行后续定量分析,通过RT-PCR,微阵列,或RNA测序。©2015 by John Wiley &儿子,Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。