期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 92, 期 7, 页码 934-942出版社
JOHN WILEY & SONS INC
DOI: 10.1002/bit.20665
关键词
DNA immobilization; hydrogel; confocal microscopy; DNA hybridization
Using a non-contact microarrayer, amine-terminated probe oligonucleotides representing 20-, 50-, and 70-mer fragments of the fliC gene were covalently coupled into three-dimensional regions in a sugar polyacrylate hydrogel based on poly(6-acryloyl-beta-O-methyl galactopyranoside-co-aminopropyl methacrylamide). The arrayer deposited the solution containing ssDNA probes in discrete regions on the surface of the gel (i.e. as a droplet with a ca. 450 mu m diameter), allowing penetration and attachment of the ss DNA within the three dimensional region of the gel. The attachment was mediated by the homobifunctional crosslinker bis-succinimidyl suberate. Confocal microscopy showed the density of attached probe DNA was greatest in the interior-most regions of the gel volume. Target ssDNA (20- and 70-mer) was able to diffuse through the gel and undergo successful hybridization with the probes. For target ssDNA in the concentration range 0.19 mu M to 6.0 mu M, there was a linear correlation between DNA concentration and the fluorescence of the gel region where hybridization occurred. (c) 2005 Wiley Periodicals, Inc.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据