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Title: Integrated Bare Narrow Capillary-Hydrodynamic Chromatographic System for Free-Solution DNA Separation at the Single-Molecule Level

Abstract

We have recently introduced a new technique, Bare Narrow Capillary Hydrodynamic Chromatography (BaNC–HDC), to resolve DNA without sieving matrices. In this technique, the separation capillaries had a diameter usually from a few hundred nanometers to a few micrometers. While high resolutions were obtained (especially for large DNA fragments), the sample injection and DNA elution were tedious, and the separation results (peak areas) were not reproducible. This was caused by the extremely low injection volumes (a few picoliters) and elution rates (<1 nanoliter per minute), which prevented us from utilizing reliable injection valves and commercial high performance liquid chromatographic pumps. In this work, we incorporate a microfabricated chip injector, and a high-pressure electroosmotic pump (EOP) to address these issues. The chip injector allows us to reliably and reproducibly inject low volumes (as low as ~1 pL) of samples into the narrow separation capillary, while the EOP provides flow rates and pressures perfectly suited for BaNC–HDC separations. Using the above system, we resolve kilo base-pair DNA fragments rapidly (in a few min) with high resolutions, and the limit of detection for double stranded DNA reaches a few molecules. The experimental parameters affecting resolutions are also discussed. Incorporation of a chip injection schememore » makes automating BaNC–HDC separations readily achievable.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [2];  [1];  [1]
  1. Univ. of Oklahoma, Norman, OK (United States). Dept. of Chemistry and Biochemistry
  2. Wuhan Textile Univ., Wuhan, Hubei (United States)
Publication Date:
Research Org.:
MicroChem Solutions, Norman, OK (United States)
Sponsoring Org.:
USDOE; National Science Foundation (NSF); Oklahoma Center for the Advancement of Science and Technology
Contributing Org.:
Univ. of Oklahoma, Norman, OK (United States
OSTI Identifier:
1337181
Grant/Contract Number:  
SC0006351; CHE 1011957; AR11-003
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Angewandte Chemie (International Edition)
Additional Journal Information:
Journal Volume: 52; Journal Issue: 21; Journal ID: ISSN 1433-7851
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; chromatography; DNA separation; electroosmotic pump; integrated system; microfluidic chip

Citation Formats

Zhu, Zaifang, Chen, Huang, Wang, Wei, Morgan, Aaron, Gu, Congying, He, Chiyang, Lu, Joann J., and Liu, Shaorong. Integrated Bare Narrow Capillary-Hydrodynamic Chromatographic System for Free-Solution DNA Separation at the Single-Molecule Level. United States: N. p., 2013. Web. doi:10.1002/anie.201300208.
Zhu, Zaifang, Chen, Huang, Wang, Wei, Morgan, Aaron, Gu, Congying, He, Chiyang, Lu, Joann J., & Liu, Shaorong. Integrated Bare Narrow Capillary-Hydrodynamic Chromatographic System for Free-Solution DNA Separation at the Single-Molecule Level. United States. https://doi.org/10.1002/anie.201300208
Zhu, Zaifang, Chen, Huang, Wang, Wei, Morgan, Aaron, Gu, Congying, He, Chiyang, Lu, Joann J., and Liu, Shaorong. Mon . "Integrated Bare Narrow Capillary-Hydrodynamic Chromatographic System for Free-Solution DNA Separation at the Single-Molecule Level". United States. https://doi.org/10.1002/anie.201300208. https://www.osti.gov/servlets/purl/1337181.
@article{osti_1337181,
title = {Integrated Bare Narrow Capillary-Hydrodynamic Chromatographic System for Free-Solution DNA Separation at the Single-Molecule Level},
author = {Zhu, Zaifang and Chen, Huang and Wang, Wei and Morgan, Aaron and Gu, Congying and He, Chiyang and Lu, Joann J. and Liu, Shaorong},
abstractNote = {We have recently introduced a new technique, Bare Narrow Capillary Hydrodynamic Chromatography (BaNC–HDC), to resolve DNA without sieving matrices. In this technique, the separation capillaries had a diameter usually from a few hundred nanometers to a few micrometers. While high resolutions were obtained (especially for large DNA fragments), the sample injection and DNA elution were tedious, and the separation results (peak areas) were not reproducible. This was caused by the extremely low injection volumes (a few picoliters) and elution rates (<1 nanoliter per minute), which prevented us from utilizing reliable injection valves and commercial high performance liquid chromatographic pumps. In this work, we incorporate a microfabricated chip injector, and a high-pressure electroosmotic pump (EOP) to address these issues. The chip injector allows us to reliably and reproducibly inject low volumes (as low as ~1 pL) of samples into the narrow separation capillary, while the EOP provides flow rates and pressures perfectly suited for BaNC–HDC separations. Using the above system, we resolve kilo base-pair DNA fragments rapidly (in a few min) with high resolutions, and the limit of detection for double stranded DNA reaches a few molecules. The experimental parameters affecting resolutions are also discussed. Incorporation of a chip injection scheme makes automating BaNC–HDC separations readily achievable.},
doi = {10.1002/anie.201300208},
url = {https://www.osti.gov/biblio/1337181}, journal = {Angewandte Chemie (International Edition)},
issn = {1433-7851},
number = 21,
volume = 52,
place = {United States},
year = {2013},
month = {4}
}

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Cited by: 9 works
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