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Simple Bulk Readout of Digital Nucleic Acid Quantification Assays
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JoVE Journal Biology
Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

DOI:
10.3791/52925-v

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06:55 min

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September 24, 2015

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,
1Broad Institute, 2Department of Biological Engineering,Massachusetts Institute of Technology

Chapters

  • 00:05Title
  • 01:03Preparing Reaction Mix for Bulk Droplet Readout
  • 02:47Droplet Formation
  • 04:05Isothermal Amplification, Data Acquisition and Analysis
  • 05:12Results: Quantification of Nucleic Acids with a Simplified Readout
  • 06:33Conclusion

Summary

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Automatic Translation

We describe an endpoint digital assay for quantifying nucleic acids with a simplified (analog) readout. We measure bulk fluorescence of droplet-based digital assays using a standard qPCR machine rather than specialized instrumentation and confirm our results by microscopy.

Tags

Keywords: Digital Nucleic Acid QuantificationDigital AssaysDroplet-based Digital AssaysBulk Fluorescence ReadoutReal-time PCRDigital PCRMultiple Displacement Amplification (MDA)Whole Genome Amplification (WGA)Quantitative AssaysSimplified Readout

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