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31 August 2017 Microfluidics-based digital quantitative PCR for single-cell small RNA quantification
Tian Yu, Chong Tang, Ying Zhang, Ruirui Zhang, Wei Yan
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Abstract

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 tomiRNA 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.

Summary Sentence

We report a novel digital quantitative PCR method for single-cell small RNA analyses.

© The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
Tian Yu, Chong Tang, Ying Zhang, Ruirui Zhang, and Wei Yan "Microfluidics-based digital quantitative PCR for single-cell small RNA quantification," Biology of Reproduction 97(3), 490-496, (31 August 2017). https://doi.org/10.1093/biolre/iox102
Received: 1 May 2017; Accepted: 29 August 2017; Published: 31 August 2017
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