Dextran sulfate provides a quantitative and quick microarray hybridization reaction

化学 核酸热力学 寡核苷酸 互补DNA 动力学 微阵列 色谱法 分子生物学 生物化学 生物 基因表达 基因 核糖核酸 物理 量子力学
作者
Wei‐Chi Ku,Wai Kwan Lau,Yu-Tien Tseng,Chi-Meng Tzeng,Sung-Kay Chiu
出处
期刊:Biochemical and Biophysical Research Communications [Elsevier]
卷期号:315 (1): 30-37 被引量:26
标识
DOI:10.1016/j.bbrc.2004.01.013
摘要

Microarray technology is a powerful tool to speed up genomics study, yet many technical aspects need to be improved. The hybridization reaction of microarray experiments is carried out for 16h or overnight in order to obtain reasonably strong signals for analysis in the presence of high salt buffer, like SSC. However, the quantitative aspect of microarray hybridization has seldom been investigated. In this study, we showed that higher overall signals from hybridization were achieved in a buffer system containing dextran sulfate, which can accelerate the kinetics of reaction by increasing the local concentration of the reactants. The dextran sulfate containing hybridization solution increases the reaction 4-fold (median) for cDNA microarray and 29-fold for oligonucleotide microarray. More importantly, the solution also provides a quantitative hybridization reaction, where the hybridization signals are proportional to the abundance of transcript added. The enhancement in the kinetics of hybridization is due to both dextran sulfate and formamide present in the solution, but the effect is not due to the higher temperature used during the reaction. With a slightly longer reaction time the hybridization reaction with the solution allows the detection of hybridization signals from rare transcripts that is not possible with regular hybridization buffers. With appropriate washing, the enhancement of kinetics by the solution does not increase the background signals at all, allowing higher signal-to-noise ratios to be achieved.
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