Novel quasi‐interpenetrating network/gold nanoparticles composite matrices for DNA sequencing by CE

Abstract

In order to further improve ssDNA sequencing performances using quasi-interpenetrating network (quasi-IPN) as a matrix composed of linear polyacrylamide (LPA) with lower viscosity-average molecular mass (3.3 MDa) and poly(N,N-dimethylacrylamide) (PDMA), gold nanoparticles (GNPs) were prepared and added into this quasi-IPN to form polymer/metal composite sieving matrices. The studies of intrinsic viscosity and differential scanning calorimetry (DSC) on quasi-IPN and quasi-IPN/GNPs indicate that there were interactions between GNPs and polymer chains. The sequencing performances on ssDNA using quasi-IPN and quasi-IPN/GNPs (with different GNPs concentrations) as sieving matrices were studied and compared by CE at different temperatures. The results show that resolutions of quasi-IPN/GNPs were higher than those of quasi-IPN without GNPs and approximated those of quasi-IPN composed of LPA with higher MW (6.5 MDa) and PDMA without GNPs in the bare fused-silica capillaries. Furthermore, the sequencing time of quasi-IPN/GNPs was shorter than that of quasi-IPN under the same sequencing conditions. The influences of GNPs and sequencing temperature on the sequencing performances of ssDNA were also discussed. The separation reproducibility of quasi-IPN/GNPs solution was excellent and its shelf life was more than 8 months.

Effects of novel quasi‐interpenetrating network/gold nanoparticles composite matrices on DNA sequencing performances by CE

“…Based on our previous work [36], in this study, in order to further investigate the effects of the composite matrices on ssDNA sequencing performances and indicate the possible functions of GNPs, we prepared quasi-IPN/GNPs composite matrices in various combinations of LPA (1.5, 3.3, and 6.5 MDa) and GNPs (about 20, 40, and 60 nm) at different GNPs contents and solution concentrations, and applied them to ssDNA sequencing, respectively. The effects of the parameters in relation to quasi-IPN/GNPs composite sieving matrices, such as GNPs contents, GNPs particle sizes, LPA molecular masses, and solution concentrations, on ssDNA sequencing performances were studied in detail by CE.…”

“…In order to prepare LPA with high molecular mass, a great quantity of expensive bis(2-ethylhexyl)sulfosuccinate (AOT) was used as an emulsifier. In previous paper [36], we prepared LPA with lower molecular mass (3.3 MDa) using a small amount of cheap sorbitan monooleate (Span 80) emulsifier. To improve the ssDNA sequencing properties using quasi-IPN formed by LPA with lower molecular mass and PDMA, we added about 40 nm gold nanoparticles (GNPs) into this quasi-IPN to form polymer/metal composite matrices.…”

Effect of Nanoparticles in Protein Separation by Capillary Electrophoresis

“…A number of different polymers such as cellulose derivatives including methyl cellulose (MC), 15 hydroxyethyl cellulose (HEC), 16 and hydroxypropylmethyl cellulose (HP-MC), 17 linear polyacrylamide (LPA), 18 poly-N,N-dimethylacrylamide (PDMA), 19 polyvinyl pyrrolidone (PVP), 20 polyvinyl alcohol (PVA), 21 and polyethylene oxide (PEO) 22 have been employed. Most of the polymers require the capillary wall-coating, resulting in the problems associated with capillary fouling, coating inhomogeneity, and limited lifetime.…”

Analysis of Broad-Range DNA Fragments with Yttrium Oxide or Ytterbium Oxide Nanoparticle/Polymer Sieving Matrix Using High-Performance Capillary Electrophoresis

“…To circumvent these difficulties, an alternative dynamic coating protocol using mainly hydrogen bonding between the Si-OH group of the capillary inner wall and the water-soluble polymers such as PEO, PVP, and PDMA have been successfully applied. 16,17,19 Recent developments on DNA separation matrix have focused on the combination of good separation efficiency with dynamic coating and low viscosity for high throughput analysis. For example, mannitol added poly(N-isopropylacrylamide) (PNIPAM), 20 gold nanoparticle (GNP), 21 and synthetic PEO 7 showed the potential for this purpose.…”

“…11 The development of separation medium is one of the most important factors for DNA separation by CE since migration behavior and resolution of DNA fragments are determined by a sieving matrix. A number of different polymers such as cellulose derivatives including methyl cellulose, 12 hydroxyethyl cellulose, 13 and hydroxypropylmethyl cellulose, 14 linear polyacrylamide, 15 polyethylene oxide, 16 polyvinyl pyrrolidone, 17 polyvinyl alcohol, 18 and poly-N,N-dimethylacrylamide (PDMA) 19 had been employed. Most of the polymers require the use of the coated capillary, which basically reduces both electrosmotic flow and the adsorption of DNA onto the capillary inner wall.…”