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Publication : Spatial and Quantitative Detection of BMP Activity in Mouse Embryonic Limb Buds.

First Author  Marques MR Year  2019
Journal  Methods Mol Biol Volume  1891
Pages  201-219 PubMed ID  30414135
Mgi Jnum  J:276937 Mgi Id  MGI:6303666
Doi  10.1007/978-1-4939-8904-1_15 Citation  Marques MR, et al. (2019) Spatial and Quantitative Detection of BMP Activity in Mouse Embryonic Limb Buds. Methods Mol Biol 1891:201-219
abstractText  Modulation of bone morphogenetic protein (BMP) activity is essential to the progression of limb development in the mouse embryo. Genetic disruption of BMP signaling at various stages of limb development causes defects ranging from complete limb agenesis to oligodactyly, polydactyly, webbing, and chondrodysplasia. To probe the state of BMP signaling in early limb buds, we designed two sets of primers to measure both spatially and quantitatively the transcription of nine key genes indicative of canonical BMP activity. One set is used to generate digoxigenin (DIG)-labeled antisense RNA probes for whole-mount mRNA in situ hybridization, while the second set is used for SYBR((R)) Green-based quantitative PCR on limb bud cDNA. Here we describe step-by-step protocols for both methods around this specific set of genes.
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