Reproducibility, bioinformatic analysis and power of the SAGE method to evaluate changes in transcriptome.

Reproducibility, bioinformatic analysis and power of the SAGE method to evaluate changes in transcriptome.
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DOI:
10.1093/nar/gni025
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发表时间:
2005-02-16
影响因子:
14.9
通讯作者:
St-Amand J
St-Amand J
中科院分区:
生物学2区
文献类型:
--
作者:
Dinel S;Bolduc C;Belleau P;Boivin A;Yoshioka M;Calvo E;Piedboeuf B;Snyder EE;Labrie F;St-Amand J

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基因表达系列分析(SAGE)方法用于研究各种实验条件下细胞或组织中的全局基因表达。然而,其再现性尚未得到明确评估。在本研究中,我们评估了 SAGE 方法的重现性并确定了影响其的因素。 SAGE 重现性的决定系数 (R2) 为 0.96。然而,有一些因素会影响 SAGE 的重现性,例如串联体和双标签的复制、测序标签的数量以及双标签的双 PCR 扩增。因此,必须对这些因素进行修正,以确保 SAGE 结果的可重复性和准确性。为了消除这些伪影,还对 SAGE 数据进行了生物信息学分析。最后,当前的研究表明,增加测序标签的数量可以提高检测转录本的方法及其在实验条件下的调节能力。
The serial analysis of gene expression (SAGE) method is used to study global gene expression in cells or tissues in various experimental conditions. However, its reproducibility has not yet been definitively assessed. In this study, we have evaluated the reproducibility of the SAGE method and identified the factors that affect it. The determination coefficient (R2) for the reproducibility of SAGE is 0.96. However, there are some factors that can affect the reproducibility of SAGE, such as the replication of concatemers and ditags, the number of sequenced tags and double PCR amplification of ditags. Thus, corrections for these factors must be made to ensure the reproducibility and accuracy of SAGE results. A bioinformatic analysis of SAGE data is also presented in order to eliminate these artifacts. Finally, the current study shows that increasing the number of sequenced tags improves the power of the method to detect transcripts and their regulation by experimental conditions.
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