Journal
ANALYTICAL CHEMISTRY
Volume 78, Issue 13, Pages 4409-4415Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac060193u
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Funding
- NHGRI NIH HHS [R01 HG001970, R01 HG001970-08, 5 R01 HG-01970-02S1] Funding Source: Medline
- NIAID NIH HHS [AI-03-017] Funding Source: Medline
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We describe a novel class of DNA separation media for microchip electrophoresis, physically cross-linked block copolymer networks, which provide rapid (<4.5 min) and remarkably enhanced resolution of DNA in a size range critical for genotyping. Linear poly( acrylamide-co-dihexylacrylamide) (LPA-co-DHA) comprising as little as 0.13 mol % dihexylacrylamide yields substantially improved electrophoretic DNA separations compared to matched molar mass linear polyacrylamide. Single-molecule videomicroscopic images of DNA electrophoresis reveal novel chain dynamics in LPA-co-DHA matrixes, resembling inchworm movement, to which we attribute the increased DNA resolution. Substantial improvements in DNA peak separation are obtained, in particular, in LPA-co-DHA solutions at polymer/copolymer concentrations near the interchain entanglement threshold. Higher polymer concentrations yield enhanced separations only for small DNA molecules (< 120 base pairs). Hydrophobically cross-linked networks offer advantages over conventional linear polymers based on enhanced separation performance ( or speed) and over chemically cross-linked gels because hydrophobic cross-links can be reversibly broken, allowing facile microchannel loading.
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