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使用荧光标记的互补DNA进行微阵列玻片杂交。

Microarray slide hybridization using fluorescently labeled cDNA.

作者信息

Ares Manuel

出版信息

Cold Spring Harb Protoc. 2014 Jan 1;2014(1):124-9. doi: 10.1101/pdb.prot080135.

Abstract

Microarray hybridization is used to determine the amount and genomic origins of RNA molecules in an experimental sample. Unlabeled probe sequences for each gene or gene region are printed in an array on the surface of a slide, and fluorescently labeled cDNA derived from the RNA target is hybridized to it. This protocol describes a blocking and hybridization protocol for microarray slides. The blocking step is particular to the chemistry of "CodeLink" slides, but it serves to remind us that almost every kind of microarray has a treatment step that occurs after printing but before hybridization. We recommend making sure of the precise treatment necessary for the particular chemistry used in the slides to be hybridized because the attachment chemistries differ significantly. Hybridization is similar to northern or Southern blots, but on a much smaller scale.

摘要

微阵列杂交用于确定实验样品中RNA分子的数量和基因组来源。针对每个基因或基因区域的未标记探针序列以阵列形式印在载玻片表面,源自RNA靶标的荧光标记cDNA与之杂交。本方案描述了微阵列载玻片的封闭和杂交方案。封闭步骤特定于“CodeLink”载玻片的化学性质,但它提醒我们,几乎每种微阵列在印刷后但杂交前都有一个处理步骤。由于附着化学性质差异很大,我们建议确定用于待杂交载玻片的特定化学性质所需的精确处理方法。杂交类似于Northern或Southern印迹,但规模要小得多。

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