MicroRNAs are small noncoding but functionally important RNA molecules that are involved in regulating diverse cellular, metabolic, and immune processes. Their small size necessitates modification in traditional acid phenol-chloroform based RNA isolation procedures to get highly enriched fraction of small RNA that includes miRNAs and siRNAs. Further, of the different methods available, real-time PCR is a powerful tool for precise and specific detection and quantification of miRNA. Moreover, real-time PCR is used to validate the screening or expression of miRNAs that are discovered during high-throughput sequencing, or microarray analysis. We demonstrate here the method of extraction of miRNAs from cultured PBMCs of bubaline origin followed by the qPCR-based (both SYBR green and TaqMan-based chemistries) identification of miRNAs expressed in response to TLR ligand stimulation.
CITATION STYLE
Mukhopadhyay, C. S., Verma, R., & Singh, J. (2017). Extraction and qPCR-Based Detection of miRNAs from cultured PBMCs of bubaline origin. In Methods in Molecular Biology (Vol. 1656, pp. 89–102). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7237-1_4
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