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Nat Biotechnol. 2015 Jul;33(7):730-2. doi: 10.1038/nbt.3246. Epub 2015 Jun 22.

Kinetic fingerprinting to identify and count single nucleic acids.

Author information

1
Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, Michigan, USA.
2
Beijing National Laboratory for Molecular Sciences, MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
3
Department of Internal Medicine, Division of Hematology/Oncology, University of Michigan, Ann Arbor, Michigan, USA.
4
1] Department of Internal Medicine, Division of Hematology/Oncology, University of Michigan, Ann Arbor, Michigan, USA. [2] Department of Internal Medicine, Division of Molecular Medicine and Genetics, University of Michigan, Ann Arbor, Michigan, USA. [3] Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA. [4] Center for Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, Michigan, USA. [5] Biointerfaces Institute, University of Michigan, Ann Arbor, Michigan, USA.

Abstract

MicroRNAs (miRNAs) have emerged as promising diagnostic biomarkers. We introduce a kinetic fingerprinting approach called single-molecule recognition through equilibrium Poisson sampling (SiMREPS) for the amplification-free counting of single unlabeled miRNA molecules, which circumvents thermodynamic limits of specificity and virtually eliminates false positives. We demonstrate high-confidence, single-molecule detection of synthetic and endogenous miRNAs in both buffer and minimally treated biofluids, as well as >500-fold discrimination between single nucleotide polymorphisms.

PMID:
26098451
PMCID:
PMC4559481
DOI:
10.1038/nbt.3246
[Indexed for MEDLINE]
Free PMC Article

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