Abstract
Detecting low-abundance long noncoding RNAs (lncRNAs) is extremely difficult due to their expression levels. Deeper sequencing with extensive protocols is required to detect these RNAs and high-throughput screens to examine the regulation of these RNAs are challenging. This protocol provides a multiplexed and robust method of detecting low-abundance RNAs, with improved signal-to-noise ratio using RNAscope-based RNA-FISH which utilizes a series of amplification steps. We have validated this protocol for investigating the regulation of low-abundance lncRNAs, which would be ideal for in vitro screening in 96-well plates.
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Acknowledgments
We would like to thank Jason S. Carroll, Susan G Komen leadership grant, Light Microscopy Core Facility, Histopathology and ISH Core Facility, and Morgane Rouault (ACD/Bio-Techne).
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Jones, J., Zecchini, H., Nagarajan, S. (2020). Multiplexed Detection and Analysis of Low-Abundance Long Noncoding RNA Using RNAscope™ in Cultured Cells. In: Nielsen, B.S., Jones, J. (eds) In Situ Hybridization Protocols . Methods in Molecular Biology, vol 2148. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0623-0_7
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DOI: https://doi.org/10.1007/978-1-0716-0623-0_7
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0622-3
Online ISBN: 978-1-0716-0623-0
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