Microarray hybridization with fractionated cDNA: Enhanced identification of differentially expressed genes
Microarray hybridization with fractionated cDNA: Enhanced identification of differentially expressed genes
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DOI：
10.1006/abio.2000.4831
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发表时间：
2000-12-01
影响因子：
2.9
通讯作者：
Kato, K
中科院分区：
文献类型：
--
作者：
Sakai, K;Higuchi, H;Kato, K
分子索引是通过将接头连接到由IIS类限制性酶消化的cDNA片段来选择cDNA亚群的技术。通过该技术，通过选择性连接接头和随后的PCR扩增将3'端cDNA片段分成16个级分。每个级分用作微阵列杂交的杂交靶。这种分级分离的靶具有相对较低的核酸复杂性，包括用于稀有转录物的更多级分，并且可用于它们的检测。在小鼠小脑和大脑RNA的实验中，对候选基因子集的分析表明，通过这种方法检测到的差异表达基因是使用未分级靶点的常规方法的10倍。(C)北京大学出版社.
Molecular indexing is a technique to select a subpopulation of cDNA by ligation of adapters to cDNA fragments digested by a class IIS restriction enzyme(s). By this technique, 3' end cDNA fragments are divided into 16 fractions by selective ligation of adapters and subsequent PCR amplification Each fraction is used as a hybridization target for microarray hybridization. This fractionated target has a relatively lower nucleic acid complexity, including more fractions for rare transcripts, and is useful for their detection. In an experiment with mouse cerebellum and cerebrum RNA, analysis of a subset of candidate genes indicated that 10 times more differentially expressed genes were detected by this method than by the conventional method using the unfractionated target. (C) 2000 Academic Press.