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Curr Protoc Mol Biol. 2015 Oct 1;112:25A.3.1-23. doi: 10.1002/0471142727.mb25a03s112.

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

Author information

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.

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

PMID:
26423587
DOI:
10.1002/0471142727.mb25a03s112
[Indexed for MEDLINE]

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