Quantitative analyses of small RNAs at the single-cell level have been challenging because of limited sensitivity and specificity of conventional real-time quantitative PCR methods. A digital quantitative PCR (dqPCR) method for miRNA quantification has been developed, but it requires the use of proprietary stem-loop primers and only applies to miRNA quantification. Here, we report a microfluidics-based dqPCR (mdqPCR) method, which takes advantage of the Fluidigm BioMark HD system for both template partition and the subsequent high-throughput dqPCR. Our mdqPCR method demonstrated excellent sensitivity and reproducibility suitable for quantitative analyses of not only miRNAs but also all other small RNA species at the single-cell level. Using this method, we discovered that each sperm has a unique miRNA profile.
CITATION STYLE
Yu, T., Tang, C., Zhang, Y., Zhang, R., & Yan, W. (2017). Microfluidics-based digital quantitative PCR for single-cell small RNA quantification. Biology of Reproduction, 97(3), 490–496. https://doi.org/10.1093/biolre/iox102
Mendeley helps you to discover research relevant for your work.