Premium
Flowable networks as DNA sequencing media in capillary columns
Author(s) -
Menchen Steve,
Johnson Ben,
Winnik Mitchell A.,
Xu Bai
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170909
Subject(s) - peg ratio , capillary electrophoresis , aqueous solution , chemistry , capillary action , polymer , electrophoresis , micelle , dna , polyethylene glycol , chromatography , rheology , chemical engineering , materials science , analytical chemistry (journal) , organic chemistry , biochemistry , finance , engineering , economics , composite material
A novel class of materials that self‐assemble in water into equilibrium network structures with a well‐defined mesh size consist of polyethylene glycols (PEG's) end‐capped with micelle‐forming fluorocarbon tails. These micellar systems form flowable aqueous gel‐like networks that permit electrophoretic DNA sequencing in capillary columns. The gels have unusual rheological properties, including netword breakdown under shear, resulting in plug flow that allows colums refill with complete ejection of byproducts of the previous sequencing analysis. In this system, DNA fragment electrophoretic mobilities are unaffected by the hydrophobicity of the polymer tails. Low molecular weight ( M ) PEG chains ( M 8000) show catastrophic resolution loss for DNA fragments larger than 100 bases due to band broadening. For a longer PEG segment ( M 35000) separating the end groups, band broadening occurs for DNA fragments larger than 300 bases, implying that the PEG segment length controls the mesh size in the equilibrium network structure. Optimum sequencing results were obtained from a 6% solution of a 1:1 mixture of C 6 F 13 end‐capped‐ and C 8 F 17 end‐capped PEG 35000. The resolution limit of fluorescent‐dye‐labeled sequencing products in this formulation was 450 bases in 75 μm capillaries at 200 V/cm.