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Tissue Gene Expression Analysis Using Arrayed Normalized cDNA Libraries

机译:使用阵列标准化的cDNA文库的组织基因表达分析

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摘要

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 () and can be used by other researchers as a digital expression reference.[The sequence data described in this paper have been submitted to the EMBL data library under accession nos. –AL36537.]
机译:我们已经使用了来自两个不同小鼠胚胎阶段的60,000个cDNA克隆上的寡核苷酸指纹数据,以建立标准化的cDNA克隆集。 5,376个克隆的标准化集代表不同的簇,因此几乎在所有情况下都代表不同的基因。 cDNA克隆的插入片段通过PCR扩增并点在载玻片上。将所得阵列与从六种不同成年小鼠组织制备的mRNA探针杂交。表达谱通过分层聚类技术进行分析。我们选择放射性检测是因为它结合了鲁棒性和灵敏度,并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,可以鉴定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。获得的结果可公开获得(),并且可以供其他研究人员用作数字表达参考。[本文中描述的序列数据已提交至EMBL数据库,登录号为。 –AL36537。]

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