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Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Published on: April 14, 2015

Expanding High-Fidelity Multiplexing in Ultrasensitive Single-Molecule Protein Detection via Proximity Barcoding.

Chi-Chia Wang1,2, Emily Dorsey1, Connie Wu1,2,3

  • 1Life Sciences Institute, University of Michigan, Ann Arbor, Michigan, USA.

Angewandte Chemie (International Ed. in English)
|May 28, 2026
PubMed
Summary
This summary is machine-generated.

We developed PRO-MOSAIX, a novel digital immunoassay platform for ultrasensitive protein detection. This technology overcomes multiplexing limitations, enabling accurate measurement of multiple proteins in biological samples.

Keywords:
biomarkersdigital ELISAmultiplexingsingle moleculeultrasensitive protein detection

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Assay Development

Background:

  • The human proteome is rich in biomarkers, but low protein concentrations in biofluids challenge detection.
  • Existing ultrasensitive assays struggle with accurate, high-throughput multiplexing due to cross-reactivity and limited readout channels.

Purpose of the Study:

  • To introduce PRO-MOSAIX (PROximity-barcoded Molecular On-bead Signal Amplification for Individual MultipleXing), a platform addressing multiplexing limitations in digital immunoassays.
  • To enable ultrasensitive, accurate, and high-throughput detection of multiple proteins simultaneously.

Main Methods:

  • PRO-MOSAIX integrates single-molecule protein detection with proximity ligation to generate specific "ON" signals.
  • DNA barcoding and a single signal readout channel overcome spectral overlap limitations.
  • Mitigation of false positives from DNA signal amplification enhances multiplexing fidelity.

Main Results:

  • A 15-plex PRO-MOSAIX assay was established and validated in human plasma.
  • The assay demonstrated low femtomolar sensitivity and high measurement accuracy.
  • PRO-MOSAIX is modular and compatible with standard laboratory equipment, including flow cytometry.

Conclusions:

  • PRO-MOSAIX provides a broadly accessible tool for research and clinical laboratories.
  • The platform effectively bridges the gap between analytical sensitivity and high-order multiplexing capabilities.
  • This technology advances biomarker discovery and diagnostic research by enabling comprehensive proteomic analysis.