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Measuring differential gene expression by short read sequencing: quantitative comparison to 2-channel gene expression microarrays

机译:通过短读测序测量差异基因表达:与2通道基因表达微阵列的定量比较

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Background High-throughput cDNA synthesis and sequencing of poly(A)-enriched RNA is rapidly emerging as a technology competing to replace microarrays as a quantitative platform for measuring gene expression. Results Consequently, we compared full length cDNA sequencing to 2-channel gene expression microarrays in the context of measuring differential gene expression. Because of its comparable cost to a gene expression microarray, our study focused on the data obtainable from a single lane of an Illumina 1 G sequencer. We compared sequencing data to a highly replicated microarray experiment profiling two divergent strains of S. cerevisiae. Conclusion Using a large number of quantitative PCR (qPCR) assays, more than previous studies, we found that neither technology is decisively better at measuring differential gene expression. Further, we report sequencing results from a diploid hybrid of two strains of S. cerevisiae that indicate full length cDNA sequencing can discover heterozygosity and measure quantitative allele-specific expression simultaneously.
机译:背景技术高通量的cDNA合成和富含poly(A)的RNA的测序正在迅速发展,它是一项竞争取代微阵列作为测量基因表达的定量平台的技术。结果因此,我们在测量差异基因表达的背景下,将全长cDNA测序与2通道基因表达微阵列进行了比较。由于其成本与基因表达微阵列相当,因此我们的研究集中在可从Illumina 1 G测序仪的单个泳道获得的数据上。我们将测序数据与一个高度复制的微阵列实验进行了比较,该实验分析了酿酒酵母的两个不同菌株。结论与以前的研究相比,使用了大量的定量PCR(qPCR)分析方法时,我们发现这两种技术在测量差异基因表达上都没有决定性的优势。此外,我们报告了来自酿酒酵母的两个菌株的二倍体杂交的测序结果,表明全长cDNA测序可以发现杂合性并同时测量定量等位基因特异性表达。

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