Abstract
In situ hybridization (ISH) techniques provide important information regarding gene expression in cells and tissues. Especially, ISH details complex spatial RNA expression in highly heterogeneous tissues, such as developing and mature central nervous systems, where rare genes involved in many fundamental developmental or biological events are expressed. Although several techniques have been developed to detect low levels of RNA expression, there are still problematic issues caused by a low signal-to-noise ratio after signal amplification. RNAscope is a recently developed ISH technique with high sensitivity and low background. RNAscope utilizes a unique probe system (double Z probe) to amplify signal from rare RNAs. Additionally, the double Z probe enables a significant reduction in nonspecific signal amplification. Here we report detailed procedures of the brown-color RNAscope ISH on embryonic and adult mouse retinas.
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Acknowledgments
We acknowledge grant support from the National Institutes of Health National Eye Institute (R01EY024376 and P30EY028102) and the Hermann Eye Fund.
Conflict of Interest: The authors declare no conflicts of interest.
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Kiyama, T., Mao, CA. (2020). Ultrasensitive RNAscope In Situ Hybridization System on Embryonic and Adult Mouse Retinas. In: Mao, CA. (eds) Retinal Development. Methods in Molecular Biology, vol 2092. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0175-4_11
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DOI: https://doi.org/10.1007/978-1-0716-0175-4_11
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