Advantages of mRNA amplification for microarray analysis
Advantages of mRNA amplification for microarray analysis
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DOI：
10.2144/02334mt04
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发表时间：
2002-10-01
期刊：
影响因子：
2.7
通讯作者：
Libutti, SK
中科院分区：
文献类型：
--
作者：
Feldman, AL;Costouros, NG;Libutti, SK
cDNA微阵列的广泛应用，如细针穿刺活检和激光捕获显微切割，需要能够执行阵列与微小的起始量的RNA。虽然已经提倡扩增RNA的方法，但使用扩增材料的阵列结果的保真度尚未得到充分验证。在这里，我们证明了保留保真度的阵列使用一个或两个回合的mRNA扩增，下游实时定量PCR验证。此外，阵列数据的质量上级使用总RNA获得的数据。基于这些结果，我们建议所有基于cDNA微阵列的基因表达分析常规mRNA扩增。
Expanding applications of cDNA microarrays such as fine needle aspiration biopsy and laser capture microdissection necessitate the ability to perform arrays with minute starting amounts of RNA. While methods for amplifying RNA have been advocated, the fidelity of array results using amplified material has not been fully validated. Here we demonstrate preserved fidelity in arrays using one or two rounds of mRNA amplification, validated by downstream real-time quantitative PCR. In addition, the quality of the array data was superior to that obtained using total RNA. Based on these results, we recommend routine mRNA amplification for all cDNA microaray-based analysis of gene expression.