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實(shí)驗(yàn)方法> 生物化學(xué)技術(shù)> 化學(xué)生物學(xué)實(shí)驗(yàn)技術(shù)>Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR

Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR

關(guān)鍵詞: microarray validation factors來(lái)源: 互聯(lián)網(wǎng)

Quantitative real-time PCR (qPCR) is a commonly used validation tool for confirming gene expression results obtained from microarray analysis; however, microarray and qPCR data often result in disagreement. The current study assesses factors contributing to the correlation between these methods in five separate experiments employing two-color 60-mer oligonucleotide microarrays and qPCR using SYBR green. Overall, significant correlation was observed between microarray and qPCR results (ρ=0.708, p<0.0001, n=277) using these platforms. The contribution of factors including up — vs. down-regulation, spot intensity, ρ-value, fold-change, cycle threshold (Ct ), array averaging, tissue type, and tissue preparation was assessed. Filtering of microarray data for measures of quality (fold-change and ρ-value) proves to be the most critical factor, with significant correlations of ρτ;0.80 consistently observed when quality scores are applied.

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