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Echinobase
ECB-ART-49547
Methods Mol Biol 2021 Jan 01;2219:195-216. doi: 10.1007/978-1-0716-0974-3_12.
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Methodology for Whole Mount and Fluorescent RNA In Situ Hybridization in Echinoderms: Single, Double, and Beyond.

Perillo M , Paganos P , Spurrell M , Arnone MI , Wessel GM .


Abstract
Identifying the location of a specific RNA in a cell, tissue, or embryo is essential to understand its function. Here we use echinoderm embryos to demonstrate the power of fluorescence in situ RNA hybridizations to localize sites of specific RNA accumulation in whole mount embryo applications. We add to this technology the use of various probe-labeling technologies to colabel multiple RNAs in one application and we describe protocols for incorporating immunofluorescence approaches to maximize the information obtained in situ. We offer alternatives for these protocols and troubleshooting advice to identify steps in which the procedure may have failed. Overall, echinoderms are wonderfully suited for these technologies, and these protocols are applicable to a wide range of cells, tissues, and embryos.

PubMed ID: 33074542
PMC ID: PMC7583651
Article link: Methods Mol Biol
Grant support: [+]

Antibodies: LOC763123 Ab1 acetyl-LOC590586 Ab14

References [+] :
Andrikou, Myogenesis in the sea urchin embryo: the molecular fingerprint of the myoblast precursors. 2013, Pubmed, Echinobase