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Publication : In situ hybridization methods for mouse whole mounts and tissue sections with and without additional β-galactosidase staining.

First Author  Komatsu Y Year  2014
Journal  Methods Mol Biol Volume  1092
Pages  1-15 PubMed ID  24318810
Mgi Jnum  J:210110 Mgi Id  MGI:5569571
Doi  10.1007/978-1-60327-292-6_1 Citation  Komatsu Y, et al. (2014) In situ hybridization methods for mouse whole mounts and tissue sections with and without additional beta-galactosidase staining. Methods Mol Biol 1092:1-15
abstractText  In situ hybridization is a powerful method for detecting endogenous mRNA sequences in morphologically preserved samples. We provide in situ hybridization methods, which are specifically optimized for mouse embryonic samples as whole mounts and section tissues. Additionally, beta-Galactosidase (beta-gal) is a popular reporter for detecting the expression of endogenous or exogenous genes. We reveal that 6-chloro-3-indoxyl-beta-D-galactopyranoside (S-gal) is a more sensitive substrate for beta-gal activity than 5-bromo-4-chloro-3-indolyl-beta-D-galactoside (X-gal). S-gal is advantageous where beta-gal activity is limited including early stage mouse embryos. As a result of the increased sensitivity as well as the color compatibility of S-gal, we successfully combined beta-gal staining using S-gal with in situ hybridization using DIG-labeled probes in both whole mounts and sections.
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