Highly efficient separation of dsDNA fragments on glass chips by using an ultralow viscosity sieving matrix
✍ Scribed by Hui Wang; Jianhua Qin; Zhongpeng Dai; Li Wang; Jiling Bai; Bingcheng Lin
- Publisher
- John Wiley and Sons
- Year
- 2003
- Tongue
- English
- Weight
- 355 KB
- Volume
- 26
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
Abstract
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/__Ha__eIII DNA marker could be achieved within 4 min by using 2% HPMC‐5 with 6% glucose added. Coelectrophoresing PCR product along with ϕX174/__Ha__eIII DNA sizing marker was also demonstrated by using the ultralow viscosity HPMC‐5 solution on a glass chip.