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Digital PCR pp 459-474 | Cite as

Droplet Digital PCR for Absolute Quantification of Extracellular MicroRNAs in Plasma and Serum: Quantification of the Cancer Biomarker hsa-miR-141

  • Maria D. Giraldez
  • John R. Chevillet
  • Muneesh TewariEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1768)

Abstract

Droplet-based digital PCR provides high-precision, absolute quantification of nucleic acid target sequences with wide-ranging applications for both research and clinical diagnostic applications. Droplet-based digital PCR enables absolute quantification by counting nucleic acid molecules encapsulated in discrete, volumetrically defined water-in-oil droplet partitions. The current available systems overcome the previous lack of scalable and practical technologies for digital PCR implementation. Extracellular microRNAs in biofluids (plasma, serum, urine, cerebrospinal fluid, etc.) are promising noninvasive biomarkers in multiple diseases and different clinical settings (e.g., diagnosis, early diagnosis, prediction of recurrence, and prognosis). Here we describe a protocol that enables highly precise and reproducible absolute quantification of extracellular microRNAs using droplet digital PCR.

Key words

Digital PCR MicroRNA Biofluids Plasma Serum Absolute quantification qPCR Reproducibility 

Notes

Acknowledgements

M.D.G acknowledges initial support from a Rio Hortega Fellowship and later from a Martin Escudero Fellowship. M.T. acknowledges support from the Department of Defense Peer-Reviewed Cancer Research Program Award CA100606 and NIH US National Institutes of Health Transformative R01 grant R01DK085714.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Maria D. Giraldez
    • 2
  • John R. Chevillet
    • 6
  • Muneesh Tewari
    • 1
    • 2
    • 3
    • 4
    • 5
    Email author
  1. 1.Comprehensive Cancer CenterUniversity of MichiganAnn ArborUSA
  2. 2.Department of Internal MedicineUniversity of MichiganAnn ArborUSA
  3. 3.Department of Biomedical EngineeringUniversity of MichiganAnn ArborUSA
  4. 4.Biointerfaces InstituteUniversity of MichiganAnn ArborUSA
  5. 5.Center for Computational Medicine and BioinformaticsUniversity of MichiganAnn ArborUSA
  6. 6.Institute for Systems BiologySeattleUSA

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