A highly sensitive and specific system for large-scale gene expression profiling.
A highly sensitive and specific system for large-scale gene expression profiling.
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大规模基因表达分析的高度敏感和特异性系统。
DOI：
10.1186/1471-2164-9-9
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发表时间：
2008-01-10
期刊：
影响因子：
4.4
通讯作者：
Li, Honghua
中科院分区：
文献类型：
--
作者：
Hu, Guohong;Yang, Qifeng;Cui, Xiangfeng;Yue, Gang;Azaro, Marco A.;Wang, Hui-Yun;Li, Honghua
基于基因表达的分子网络整合技术的快速发展对提高实验系统的灵敏度和数据准确性提出了强烈的要求。为了满足这一需求，一种高通量、高特异性和高灵敏度的基因表达谱分析系统应运而生。通过使用特殊设计的引物，新系统扩增被大内含子分隔的相邻外显子中的序列，使得mRNA序列可以有效地与其他高度相关的序列（包括其基因、未加工的转录物、假基因和假基因转录物）区分开。用于微阵列检测的探针由引物扩增的两个相邻外显子中的序列组成。结合新开发的高通量多重扩增系统和高度简化的实验程序，该系统可用于在单次检测中分析> 1，000种mRNA。它也可用于极少数（n = 100）或单个细胞的基因表达谱分析。从具有相同细胞数量的重复样品以及具有少量（100）和大量（10，000）细胞的重复样品中获得了高度可重复的结果。通过比较乳腺癌细胞系MCF-7和卵巢癌细胞系NCI/ADR-RES的结果，并使用基因组DNA作为起始材料，证明了该系统的特异性。我们的方法可能会大大方便在许多重要的应用，特别是当RNA的量是有限的已知基因的组合表达的分析。
Rapid progress in the field of gene expression-based molecular network integration has generated strong demand on enhancing the sensitivity and data accuracy of experimental systems. To meet the need, a high-throughput gene profiling system of high specificity and sensitivity has been developed. By using specially designed primers, the new system amplifies sequences in neighboring exons separated by big introns so that mRNA sequences may be effectively discriminated from other highly related sequences including their genes, unprocessed transcripts, pseudogenes and pseudogene transcripts. Probes used for microarray detection consist of sequences in the two neighboring exons amplified by the primers. In conjunction with a newly developed high-throughput multiplex amplification system and highly simplified experimental procedures, the system can be used to analyze >1,000 mRNA species in a single assay. It may also be used for gene expression profiling of very few (n = 100) or single cells. Highly reproducible results were obtained from duplicate samples with the same number of cells, and from those with a small number (100) and a large number (10,000) of cells. The specificity of the system was demonstrated by comparing results from a breast cancer cell line, MCF-7, and an ovarian cancer cell line, NCI/ADR-RES, and by using genomic DNA as starting material. Our approach may greatly facilitate the analysis of combinatorial expression of known genes in many important applications, especially when the amount of RNA is limited.
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影响因子：
4.4
作者：
Hu, YH;Hines, LM;LaBaer, J
通讯作者：
LaBaer, J
影响因子：
56.9
作者：
Golub, TR;Slonim, DK;Lander, ES
通讯作者：
Lander, ES
影响因子：
9.4
作者：
Aguilar, JC;Pérez-Breña, MP;Echevarria, JE
通讯作者：
Echevarria, JE
DOI：
10.1097/00001703-200402000-00010
发表时间：
2004-02-01
影响因子：
2.1
作者：
Coradini, D;Daidone, MG
通讯作者：
Daidone, MG
影响因子：
7
作者：
Pastinen, T;Raitio, M;Syvänen, AC
通讯作者：
Syvänen, AC