Chandran Divya, Scanlon Michael J, Ohtsu Kazuhiro, Timmermans Marja C P, Schnable Patrick S, Wildermuth Mary C
University of California, Berkeley, California.
Regional Center for Biotechnology, Faridabad, India.
Curr Protoc Mol Biol. 2015 Oct 1;112:25A.3.1-25A.3.23. doi: 10.1002/0471142727.mb25a03s112.
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.
对细胞进行激光显微切割能够分离出特定的目标细胞，用于包括转录谱分析在内的下游分析。由于植物细胞具有纤维素细胞壁和大液泡，因此对其进行激光显微切割存在独特的挑战。在此，我们介绍植物组织制备、选定植物细胞的激光显微切割以及从切割细胞中进行RNA线性扩增的方法。从切割细胞中进行RNA线性扩增可获得足够的RNA，用于后续通过逆转录聚合酶链反应（RT-PCR）、微阵列或RNA测序进行定量分析。
