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Related Experiment Video

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Simultaneously sizing and quantitating zeptomole-level DNA at high throughput in free solution.

Zaifang Zhu1, Huang Chen, Apeng Chen

  • 1Department of Chemistry and Biochemistry, University of Oklahoma, 101 Stephenson Parkway, Norman, OK 73019-5251 (USA).

Chemistry (Weinheim an Der Bergstrasse, Germany)
|September 17, 2014
PubMed
Summary

A new method, bare-narrow-capillary/hydrodynamic-chromatography (BaNC-HDC), accurately sizes and quantitates DNA at extremely low concentrations. This technique offers high-throughput analysis for DNA fragments from base pairs to kilobase pairs.

Keywords:
DNA sizinggel-free DNA separationhigh-throughput screeninghydrodynamic chromatographymicrochips

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Accurate DNA sizing and quantitation are crucial in molecular biology.
  • Traditional gel electrophoresis struggles to simultaneously size and quantify DNA at low zeptomole (zmol) concentrations.
  • There is a need for sensitive, high-throughput methods for DNA analysis.

Purpose of the Study:

  • To introduce and validate a novel technique, bare-narrow-capillary/hydrodynamic-chromatography (BaNC-HDC), for DNA analysis.
  • To demonstrate BaNC-HDC's capability in simultaneously sizing and quantitating DNA fragments at extremely low concentrations.
  • To assess the throughput and accuracy of BaNC-HDC for diverse DNA sizes.

Main Methods:

  • Development of bare-narrow-capillary/hydrodynamic-chromatography (BaNC-HDC) without sieving matrices.
  • Application of BaNC-HDC for resolving DNA fragments based on size.
  • Quantitation of DNA molecules at zeptomole (zmol) to several-molecule levels.

Main Results:

  • BaNC-HDC accurately sizes and quantitates DNA fragments ranging from a few base pairs to dozens of kilobase pairs.
  • The method achieves a throughput of 15 samples per hour, analyzing samples with multiple DNA lengths.
  • Successful detection and quantification of DNA at zeptomole (zmol) to single-molecule concentrations were demonstrated.

Conclusions:

  • BaNC-HDC is a powerful tool for high-throughput DNA sizing and quantitation at very low concentrations.
  • This matrix-free chromatography offers a cost-effective alternative to existing methods.
  • BaNC-HDC significantly advances the ability to analyze minute quantities of DNA in molecular biology research.