Laser Microdissection–Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA. Issue 1 (15th February 2018)

• Record Type: Journal Article
• Authors: Chandran, Divya; Scanlon, Michael J.; Ohtsu, Kazuhiro; Timmermans, Marja C.P.; Schnable, Patrick S.; Wildermuth, Mary C.
• Editors: Ausubel, Frederick M.; Brent, Roger; Kingston, Robert E.; Moore, David D.; Seidman, J.G.; Smith, John A.; Struhl, Kevin
• Is Part Of: Current protocols in molecular biology. Volume 112:Issue 1(2015)
• Page Start: 25A.3.1
• Page End: 25A.3.23
• Publication Date: 2018-02-15
• Subjects: laser microdissection -- plants -- RNA amplification -- transcriptomics; Molecular biology -- Technique -- Periodicals; Molecular biology -- Laboratory manuals; Molecular Biology -- methods; Biologie moléculaire -- Technique; Biologie moléculaire -- Manuels de laboratoire; Molecular biology; Molecular biology -- Technique; Laboratory Manual; Electronic reference sources; Laboratory manuals; 572.8028
• DOI: 10.1002/0471142727.mb25a03s112
• Languages: English
• ISSNs: 1934-3639

Abstract

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.