The specific aim of the renewal proposal deals with further development and optimization of new and better separation media which can overcome many of the problems related to the present-day polyacrylamide solutions or gels: polymer solutions are often too viscous to be filled in capillary tubings; the polymerization in situ approach is not so reproducible; shelf-life for polymer gels is limited. The key scheme is to take advantage of the self-assembly behavior of copolymer colloids. The investigators have achieved the initial goals of designing a polymer network based on the self-assembly behavior of a EPE-type block copolymer (E and P denoted polyoxyethylene and polyoxypropylene, respectively) with the following properties: (1) at 10-15 degrees C, the polymer solution is a low viscosity fluid (less than 30 centipoise) in 1xTBE buffer, for ease of filling and removing the separation medium in narrow-bore capillary tubings; (2) at room temperatures (greater than 20 degrees C), the polymer solution becomes gel-like with good sieving ability (tested down to 14 bases with one base resolution for single stranded (ss) DNA fragments and up to 1 kilobase pairs for ds DNA fragments with one base pair resolutions); and (3) the polymer itself has acted successfully as a dynamic coating material. By correlating the capillary electrophoresis results with structural studies using a variety of physical techniques, such as laser light scattering, small angle X-ray scattering, small angle neutron scattering, NMR, rheometry, viscosimetry and atomic force microscopy, an optimization of the self-assembled polymer network for DNA capillary electrophoresis can be achieved. In addition to the EPE, PEP, EBE, and BEB type triblocks (with B being polyoxybutylene and more hydrophobic than P), it is proposed to use crystallization and coil-collapse as the physical phenomena capable of temperature-dependent reversible cross-linking formation. The crystallization approach is especially attractive, since the supramolecular structure can be more resilient and is not in dynamic equilibrium with single polymer chains as in the self-assembly process.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
5R01HG001386-06
Application #
6181625
Study Section
Genome Study Section (GNM)
Program Officer
Schloss, Jeffery
Project Start
1995-09-30
Project End
2001-08-31
Budget Start
2000-09-01
Budget End
2001-08-31
Support Year
6
Fiscal Year
2000
Total Cost
$187,648
Indirect Cost
Name
State University New York Stony Brook
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Wan, Fen; He, Weidong; Zhang, Jun et al. (2009) Reduced matrix viscosity in DNA sequencing by CE and microchip electrophoresis using a novel thermo-responsive copolymer. Electrophoresis 30:2488-98
Wan, Fen; Zhang, Jun; Lau, Angela et al. (2008) Nanostructured copolymer gels for dsDNA separation by CE. Electrophoresis 29:4704-13
Zhang, Jun; He, Weidong; Liang, Dehai et al. (2006) Designing polymer matrix for microchip-based double-stranded DNA capillary electrophoresis. J Chromatogr A 1117:219-27
Wan, Fen; He, Weidong; Zhang, Jun et al. (2006) Scale-up development of high-performance polymer matrix for DNA sequencing analysis. Electrophoresis 27:3712-23
Zhang, Jun; Gassmann, Marcus; He, Weidong et al. (2006) Reversible thermo-responsive sieving matrix for oligonucleotide separation. Lab Chip 6:526-33
Zhang, Jun; Burger, Christian; Chu, Benjamin (2006) Nanostructured polymer matrix for oligonucleotide separation. Electrophoresis 27:3391-8
Zhang, Jun; Liang, Dehai; He, Weidong et al. (2005) Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen as fluorescence inducing agent. Electrophoresis 26:4449-55
Wang, Yanmei; Liang, Dehai; Ying, Qicong et al. (2005) Quasi-interpenetrating network formed by polyacrylamide and poly(N,N-dimethylacrylamide) used in high-performance DNA sequencing analysis by capillary electrophoresis. Electrophoresis 26:126-36
Liang, Dehai; Zhang, Jun; Chu, Benjamin (2003) Study of ethidium bromide effect on dsDNA separation by capillary zone electrophoresis and laser light scattering. Electrophoresis 24:3348-55
Moritani, Tohei; Yoon, Kyunghwan; Rafailovich, Miriam et al. (2003) DNA capillary electrophoresis using poly(vinyl alcohol). I. Inner capillary coating. Electrophoresis 24:2764-71

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