Novel hydroxyethylcellulose‐graft‐poly acrylamide copolymer for separation of double‐stranded DNA fragments by CE

Abstract

A novel separation medium, hydroxyethylcellulose-graft-polyacrylamide (HEC-g-PAM) synthesized by atom transfer radical polymerization (ATRP), used for dsDNA separation by CE is presented. The separation performance of HEC-g-PAM, which has the same graft density and different graft length, has been investigated in Tris-boric acid-EDTA (TBE) buffer solvent mixtures. The temperature-dependent rheological behavior of HEC-g-PAM was also studied by steady-shear rheometry. The results showed that dsDNA fragments between 72 and 1353 bp was achieved with a 30 cm effective capillary length at 150 V/cm using this type of graft copolymer as a separation medium in bare fused-silica capillaries, and separation improvement is obtained in HEC-g-PAM compared with HEC and poly(dimethylacrylamide (PDMA).

Hydroxyethylcellulose‐graft‐poly (N,N‐dimethylacrylamide) copolymer as a multifunctional separation medium for CE

“…3 since R s could be related directly to the size difference of the two DNA fragments (DB), R s /DB were normalized for 1-bp difference and used for a proper comparison of different fragment doublets. For selectivity and resolution per base pair, there was a similar tendency as the previous work of our group [3]. For shorter DNA fragments (below 300 bp), high concentration of HEC-g-PDMA has a higher resolution than low concentration.…”

“…Moreover, separation medium, which is particularly devised for the special separation purpose, possesses its unique characteristics. Taking respective separation of dsDNA and basic proteins by CE for an example, an ideal separation medium for dsDNA separations should possess the following properties: high sieving ability, dynamic coating ability, and relatively low viscosity [3]. However, the responsibility of basic protein separation medium is to minimize the protein-capillary wall interactions and suppress or stabilize EOF [4].…”

Synthesis of hydroxyethylcellulose‐graft‐poly(N, N‐dimethylacrylamide) copolymer by ATRP and as dynamic coating in capillary electrophoresis

“…Our group also successive synthesized hydroxyethylcellulose-graft-polyacrylamide by using ATRP. 7 Although both N, N-dimethylacrylamide (DMA) and PDMA are strong coordinating ligands that can compete for the binding of the metal catalysts in polymerization system, PDMA can be successfully polymerized with the molecular weight distributions. [8][9][10][11] The ligand is very important for controlled radical polymerization of DMA.…”

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ABSTRACT: Hydroxyethylcellulose-graft-poly (N, Ndimethylacrylamide) was synthesized by successive atom transfer radical polymerization (ATRP) of N,N-dimethylacrylamide (DMA) monomer using HEC-Br as initiator, CuBr and 5,5,7,12,12,4,8, [14]aneN 4 ) as catalyst and ligand, with molar ratio DMA: HEC-Br (CABr): CuBr: Me 6 [14]aneN 4 ¼ 100 : 1 : 1 : 3. HEC-Br macroinitiator was synthesized by esterification of HEC with 2-bromoisobutyryl bromide.

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Effect of quasi‐interpenetrating network of polyacrylamide and poly(N,N‐dimethylacrylamide) on separation of dsDNA fragments and basic protein using CE

“…Sieving matrices are very important for the separation and sequencing of DNA, because they determine the migration behavior and the resolution of DNA during separation [3 -5]. An ideal separation medium for dsDNA separation should possess the following properties: high sieving ability, dynamic coating ability, and relatively low viscosity [6]. A variety of polymers for DNA separation by CE have been reported in our previous review [7].…”