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使用阵列式标准化cDNA文库进行组织基因表达分析。

Tissue gene expression analysis using arrayed normalized cDNA libraries.

作者信息

Eickhoff H, Schuchhardt J, Ivanov I, Meier-Ewert S, O'Brien J, Malik A, Tandon N, Wolski E W, Rohlfs E, Nyarsik L, Reinhardt R, Nietfeld W, Lehrach H

机构信息

Max-Planck-Institut für Molekulare Genetik, 14195 Berlin, Germany.

出版信息

Genome Res. 2000 Aug;10(8):1230-40. doi: 10.1101/gr.10.8.1230.

Abstract

We have used oligonucleotide-fingerprinting data on 60,000 cDNA clones from two different mouse embryonic stages to establish a normalized cDNA clone set. The normalized set of 5,376 clones represents different clusters and therefore, in almost all cases, different genes. The inserts of the cDNA clones were amplified by PCR and spotted on glass slides. The resulting arrays were hybridized with mRNA probes prepared from six different adult mouse tissues. Expression profiles were analyzed by hierarchical clustering techniques. We have chosen radioactive detection because it combines robustness with sensitivity and allows the comparison of multiple normalized experiments. Sensitive detection combined with highly effective clustering algorithms allowed the identification of tissue-specific expression profiles and the detection of genes specifically expressed in the tissues investigated. The obtained results are publicly available (http://www.rzpd.de) and can be used by other researchers as a digital expression reference.

摘要

我们利用来自两个不同小鼠胚胎阶段的60,000个cDNA克隆的寡核苷酸指纹数据,建立了一个标准化的cDNA克隆集。这个包含5376个克隆的标准化集代表了不同的簇,因此在几乎所有情况下都代表了不同的基因。cDNA克隆的插入片段通过PCR扩增,然后点样到载玻片上。所得阵列与从六种不同成年小鼠组织制备的mRNA探针进行杂交。通过层次聚类技术分析表达谱。我们选择放射性检测是因为它兼具稳健性和灵敏度,并且允许对多个标准化实验进行比较。灵敏的检测与高效的聚类算法相结合,使得我们能够识别组织特异性表达谱,并检测在所研究组织中特异性表达的基因。所获得的结果可公开获取(http://www.rzpd.de),可供其他研究人员用作数字表达参考。

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引用本文的文献

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Normalization strategies for cDNA microarrays.cDNA微阵列的标准化策略。
Nucleic Acids Res. 2000 May 15;28(10):E47. doi: 10.1093/nar/28.10.e47.
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High density synthetic oligonucleotide arrays.高密度合成寡核苷酸阵列
Nat Genet. 1999 Jan;21(1 Suppl):20-4. doi: 10.1038/4447.
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Cluster analysis and display of genome-wide expression patterns.全基因组表达模式的聚类分析与展示
Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14863-8. doi: 10.1073/pnas.95.25.14863.

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