Highly efficient separation of dsDNA fragments on glass chips by using an ultralow viscosity sieving matrix

Abstract

Highly efficient separation of dsDNA fragments on glass chips by using an ultralow viscosity sieving matrix A novel protocol has been established to separate dsDNA fragments with high efficiency on glass chips by using an ultralow viscosity sieving matrix with added glucose. Low-molecular-weight hydroxypropylmethylcellulose (HPMC), with a viscosity nearly equivalent to that of water, was used to electrophoretically separate fluorescent intercalator-labeled double-stranded DNA (dsDNA) fragments on microfluidic glass chips. In comparison with conventional sieving protocols, low-molecular-weight HPMC as sieving matrix could result in reduced running cost and analysis time, in addition to a comparable separation efficiency of dsDNA fragments. In this paper, the addition of glucose was investigated to enhance the separation of DNA in the lowest viscosity polymer evaluated. The effect of staining dye and field strength were also evaluated. At an applied electric field strength of 200 V/cm, satisfactory resolution of the PBR322/HaeIII DNA marker could be achieved within 4 min by using 2% HPMC-5 with 6% glucose added. Coelectrophoresing PCR product along with bX174/HaeIII DNA sizing marker was also demonstrated by using the ultralow viscosity HPMC-5 solution on a glass chip.

Microchip capillary electrophoresis with laser‐induced fluorescence combined with one‐step duplex reverse‐transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens

“…With the development of microelectromechanical systems and micromachine technology, the research of MCE as an emerging separation technique has gained great attention in clinical medicine and public health surveillance . MCE offers a novel way to achieve rapid and cost‐effective separation with high resolution and low sample consumption on a miniaturized platform, which has become a promising analytical technique and been playing an increasingly important role in the detection of pathogenic viruses and bacteria .…”

Modification of poly(methyl methacrylate) microchannels for highly efficient and reproducible electrophoretic separations of double-stranded DNA

“…Microchips fabricated in a variety of substrates, including glass, quartz, and plastics, have been used for electrophoretic separations of DNA [13][14][15][16][17][18]. Polymer substrates such as polycarbonate [17] and poly(methyl methacrylate) (PMMA) [18] are particularly interesting because they offer a greater potential for making disposable devices on a cost-effective basis by printing, laser ablation, and plasma etching [18][19][20][21].…”