In situ hybridization to mRNA of Arabidopsis tissue sections

Methods : a Companion to Methods in Enzymology
J de Almeida EnglerG Engler

Abstract

In situ hybridization detection of mRNA is an essential tool for understanding regulation of gene expression in cells and tissues of different organisms. Over the years, numerous in situ protocols have been developed ranging from whole-mount techniques that allow fast transcript localization in intact organs to high-resolution methods based on the electron microscopic detection of mRNAs at the subcellular level. Here, we present a detailed protocol for the detection of mRNAs in plant tissues using radiolabeled single-stranded RNA probes. Hybridizations are carried out on tissue sections of paraffin- and plastic-embedded plant tissues. Although this in situ protocol is appropriate for plant tissues in general, it has been optimized for Arabidopsis thaliana. Variations on the procedure, required to obtain optimal results with different Arabidopsis tissues, are described.

References

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Citations

May 25, 2002·Molecular Plant-microbe Interactions : MPMI·Isabel VercauterenGodelieve Gheysen
Dec 8, 2004·Plant Physiology·Rosa Maria BarrôcoGilbert Engler
Dec 23, 2008·Plant Physiology·Adriano SenatoreJohn S Greenwood
Oct 9, 2002·The Plant Cell·Kristiina HimanenTom Beeckman
Sep 3, 2004·The Plant Journal : for Cell and Molecular Biology·Edouard PesquetDeborah Goffner
Apr 1, 2004·The Plant Journal : for Cell and Molecular Biology·Janice de Almeida EnglerGodelieve Gheysen
Apr 28, 2009·The Plant Journal : for Cell and Molecular Biology·Janice de Almeida EnglerGilbert Engler
May 30, 2012·The Plant Journal : for Cell and Molecular Biology·Janice de Almeida EnglerGodelieve Gheysen
Apr 24, 2001·Methods : a Companion to Methods in Enzymology·T W Abraham
Apr 24, 2001·Methods : a Companion to Methods in Enzymology·B K Micales, G E Lyons
Jul 28, 2016·Frontiers in Plant Science·Fernando E Díaz-ManzanoCarolina Escobar
Feb 1, 2014·Progress in Lipid Research·Patrick J Horn, Kent D Chapman
Nov 16, 2001·Phytochemical Analysis : PCA
Nov 8, 2020·International Journal of Molecular Sciences·Vishvanathan MarthandanJegadeesan Ramalingam
Mar 2, 2021·Frontiers in Plant Science·Luiz M CabralAdriana S Hemerly
May 18, 2021·Frontiers in Plant Science·Danila CabralJanice de Almeida Engler

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