Concordance among digital gene expression, microarrays, and qPCR when measuring differential expression of microRNAs
Concordance among digital gene expression, microarrays, and qPCR when measuring differential expression of microRNAs
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DOI：
10.2144/000113367
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发表时间：
2010-03-01
期刊：
影响因子：
2.7
通讯作者：
Harshman, Keith
中科院分区：
文献类型：
--
作者：
Pradervand, Sylvain;Weber, Johann;Harshman, Keith
鉴于miRNA在许多细胞功能中的关键作用，分析microRNA（miRNA）表达受到广泛关注。可以通过基于杂交（微阵列）、基于测序或基于扩增（定量逆转录-PCR，qPCR）的技术实现分析。其中，微阵列面临着准确区分成熟和未成熟miRNA形式的重大挑战，不同的供应商已经开发了不同的方法来应对这一挑战。在这里，我们使用Affytek，Agilent和Illumina微阵列平台以及qPCR（Applied Biosystems）和超高通量测序（Illumina）测量差异miRNA表达。我们发现，差异表达测量是更发散的三种类型的微阵列相比，当安捷伦微阵列，qPCR和测序技术的测量进行比较，表现出良好的整体一致性。
Profiling microRNA (miRNA) expression is of widespread interest given the critical role of miRNAs in many cellular functions. Profiling can be achieved via hybridization-based (microarrays), sequencing-based, or amplification-based (quantitative reverse transcription-PCR, qPCR) technologies. Among these, microarrays face the significant challenge of accurately distinguishing between mature and immature miRNA forms, and different vendors have developed different methods to meet this challenge. Here we measure differential miRNA expression using the Affymetrix, Agilent, and Illumina microarray platforms, as well as qPCR (Applied Biosystems) and ultra high-throughput sequencing (Illumina). We show that the differential expression measurements are more divergent when the three types of microarrays are compared than when the Agilent microarray, qPCR, and sequencing technology measurements are compared, which exhibit a good overall concordance.