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Publication : An economical and practical method for whole-mount in situ hybridization for mouse embryos and organs.

First Author  Pu D Year  2013
Journal  Biotech Histochem Volume  88
Issue  1 Pages  27-37
PubMed ID  23088431 Mgi Jnum  J:211545
Mgi Id  MGI:5575635 Doi  10.3109/10520295.2012.724712
Citation  Pu D, et al. (2013) An economical and practical method for whole-mount in situ hybridization for mouse embryos and organs. Biotech Histochem 88(1):27-37
abstractText  Whole-mount in situ hybridization (WISH) is a useful method for detecting specific gene expression patterns at their site of action during embryonic development. Traditional WISH methods are costly and suitable only for mouse embryos younger than 11.5 days. We present here an economical and practical in situ hybridization method using DIG-labeled RNA probes. We changed the conditions in several steps to make the WISH method suitable for whole mouse embryos from embryonic days 9.5 to 12.5 and for older stage mouse embryonic organs. We performed all steps in one microcentrifuge tube up to the staining steps to avoid losing or damaging the mouse embryos. We re-used the solutions and materials to make the method more economical and suitable for less sophisticated laboratories. We also performed beta-galactosidase staining on Tb x 18 Cre/Rosa26/LacZ mouse embryos; the results agreed with the in situ hybridization results. Finally, we sectioned the specimens after hybridization and beta-galactosidase staining; the results agreed with the literature.
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