With support from the Chemical Measurement and Imaging Program, Prof. Shaorong Liu and his group at the University of Oklahoma are seeking better understanding of how a narrow bore capillary can be used to effectively separate DNA fragments (from 75 to 106,000 base pairs) and proteins with efficiencies comparable to those achieved with capillary zone electrophoresis. The work entails systematic investigation to elucidate the separation mechanisms and their relative contributions.

Development of separation techniques using narrow capillaries will extend the limits of separation science - critical to research in biological, biotechnological, chemical, pharmaceutical, environmental, geochemical, and agricultural disciplines. The inherently interdisciplinary nature of this research requires new modes of student training and new curricula incorporating aspects of theory, computation, nanofluidics, and separations. Substantial education and recruitment activities in nanoscience and technology are particularly aimed at groups underrepresented in science and engineering, including recruitment through a regional network of primarily undergraduate institutions, many of which have large populations strong in underrepresented talent. Results will be incorporated into general-public and high-school level nanoscience seminar series being developed by the PI in conjunction with activists at the Norman library.

Project Report

Scientific Findings as Outcomes 1. Pressure-Induced Transport of DNA Confined in Narrow Capillary Channels. We use a hydrodynamic chromatographic (HDC) technique to study the fundamental mechanisms of pressure-induced transport of DNA ranging from 10 bp to 1.9 Mbp confined in a 750-nm-radius bare capillary. On the basis of varying transport behavior of DNA confined in such a narrow capillary, we have identified four distinct size regions. Utilizing an HDC quadratic model, we have studied the transport behavior and found proper equations (models) to estimate the mobility of DNA in all these regions. 2. Resolving DNA at High-Speed and High-Throughput without Sieving Matrices. We demonstrate the feasibility of utilizing BaNC-HDC for high-throughput sizing and quantitation of DNA at zepto-mole to several-molecule levels using a single open capillary column without any sieving matrices or wall coatings. Both sizing and quantitation accuracies are excellent (errors are generally at the single-digit-percentage level), and sample throughputs of more than a dozen have been achieved. Additional advantages of BaNC-HDC are the low waste generation (pL/min effluent to waste), the low consumable costs (the expenses of a few nL eluent and a few pL sample per assay), and readiness for automation. Peer-Reviewed Publications as Outcomes 1) Huang Chen, Zaifang Zhu, Joann J. Lu, and Shaorong Liu, On-Line Labeling and Sizing of PCR-Amplified DNA by Bare Narrow Capillary – Hydrodynamic Chromatography, 2014, Submitted. 2) Zaifang Zhu, Apeng Chen, Huang Chen, Joann J. Lu, and Shaorong Liu, Resolving DNA at High-Speed and High-Throuput without Sieving Matrices, Chemistry – A European Journal, 2014, accepted as a cover-page article (http://onlinelibrary.wiley.com/doi/10.1002/chem.201404998/pdf) 3) Zaifang Zhu, Huang Chen, Wei Wang, Aaron Morgan, Congying Gu, Chiyang He, Joann J. Lu, and Shaorong Liu, Integrated Bare Narrow Capillary–Hydrodynamic Chromatographic System for Free-Solution DNA Separation at Single-Molecule Level, Angew. Chem. Int. Ed. 52 (2013), 5612 –5616. 4) Zaifang Zhu, Lei Liu, Wei Wang, Joann J. Lu, Xiayan Wang and Shaorong Liu, Resolving DNA at efficiencies of more than a million plates per meter using bare narrow open capillaries without sieving matrices, Chem. Commun., 49 (2013), 2897–2899. 5) Xiayan Wang, Lei Liu, Qiaosheng Pu, Guangsheng Guo, Hui Zhong and Shaorong Liu, Pressure-Induced Transport of DNA Confined in Narrow Capillary Channels, J. Am. Chem. Soc., 134 (2012), 7400–7405. 6) Xiayan Wang, Lei Liu, Wei Wang, Qiaosheng Pu, Guangsheng Guo, Purnendu K. Dasgupta and Shaorong, Liu, Resolving DNA in free solution, Trends Anal. Chem. 35 (2012), 122-134. 7) Xiayan Wang, Vijaykumar Veerappan, Chang Cheng, Xin Jiang, Randy D. Allen, Purnendu K. Dasgupta, and Shaorong Liu, Free Solution Hydrodynamic Separation of DNA Fragments from 75 to 106000 Base Pairs in A Single Run, J. Am. Chem. Soc., 132 (2010), 40–41. Personell Trainging as outcomes Current Graduate Student: Huang Chen, Ph.D program Ph.D Students Graduated: Zaifang Zhu, Accepted a Faculty Position at Lanzhou University, Lanzhou, P.R.China Postdoctoral Fellow Trained: Xiayan Wang, Now Working as a Full Professor at Beijing University of Technology, Beijing, P.R.China

Agency
National Science Foundation (NSF)
Institute
Division of Chemistry (CHE)
Type
Standard Grant (Standard)
Application #
1011957
Program Officer
Kelsey Cook
Project Start
Project End
Budget Start
2010-09-01
Budget End
2014-08-31
Support Year
Fiscal Year
2010
Total Cost
$375,000
Indirect Cost
Name
University of Oklahoma
Department
Type
DUNS #
City
Norman
State
OK
Country
United States
Zip Code
73019