Laser Microdissection-Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA

Divya Chandran; Michael J Scanlon; Kazuhiro Ohtsu; Marja C P Timmermans; Patrick S Schnable; Mary C Wildermuth

doi:10.1002/0471142727.mb25a03s112

Laser Microdissection-Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA

Curr Protoc Mol Biol. 2015 Oct 1:112:25A.3.1-25A.3.23. doi: 10.1002/0471142727.mb25a03s112.

Authors

Divya Chandran^{1

2}, Michael J Scanlon³, Kazuhiro Ohtsu⁴, Marja C P Timmermans⁵, Patrick S Schnable⁴, Mary C Wildermuth¹

Affiliations

¹ University of California, Berkeley, California.
² Regional Center for Biotechnology, Faridabad, India.
³ Cornell University, Ithaca, New York.
⁴ Iowa State University, Ames, Iowa.
⁵ Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

PMID: 26423587
DOI: 10.1002/0471142727.mb25a03s112

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.

Keywords: RNA amplification; laser microdissection; plants; transcriptomics.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Laser Capture Microdissection / methods*
Nucleic Acid Amplification Techniques / methods*
Plant Cells*
RNA, Plant / genetics*
RNA, Plant / isolation & purification*

Substances

RNA, Plant