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Updated: Feb 2, 2026

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR
Published on: May 29, 2014
Spatial and Quantitative Detection of BMP Activity in Mouse Embryonic Limb Buds
Marcelo Rocha Marques1,2, Jean-Denis Bénazet3
1Department of Morphology, Area of Histology and Embryology, Piracicaba Dental School, University of Campinas, Campinas, SP, Brazil.
Abstract:
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® 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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