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用于全基因组表达研究的区域性解剖人类对照大脑的大型数据集的质量控制参数。

Quality control parameters on a large dataset of regionally dissected human control brains for whole genome expression studies.

机构信息

Reta Lilla Weston Laboratories and Departments of Molecular Neuroscience, UCL Institute of Neurology, London, UK.

出版信息

J Neurochem. 2011 Oct;119(2):275-82. doi: 10.1111/j.1471-4159.2011.07432.x. Epub 2011 Sep 20.

Abstract

We are building an open-access database of regional human brain expression designed to allow the genome-wide assessment of genetic variability on expression. Array and RNA sequencing technologies make assessment of genome-wide expression possible. Human brain tissue is a challenging source for this work because it can only be obtained several and variable hours post-mortem and after varying agonal states. These variables alter RNA integrity in a complex manner. In this report, we assess the effect of post-mortem delay, agonal state and age on gene expression, and the utility of pH and RNA integrity number as predictors of gene expression as measured on 1266 Affymetrix Exon Arrays. We assessed the accuracy of the array data using QuantiGene, as an independent non-PCR-based method. These quality control parameters will allow database users to assess data accuracy. We report that within the parameters of this study post-mortem delay, agonal state and age have little impact on array quality, array data are robust to variable RNA integrity, and brain pH has only a small effect on array performance. QuantiGene gave very similar expression profiles as array data. This study is the first step in our initiative to make human, regional brain expression freely available.

摘要

我们正在构建一个开放获取的区域人类大脑表达数据库,旨在允许对表达进行全基因组评估的遗传变异性。阵列和 RNA 测序技术使全基因组表达的评估成为可能。人脑组织是这项工作的一个具有挑战性的来源,因为它只能在死后几个小时到不同的濒死状态和不同的年龄获得。这些变量以复杂的方式改变 RNA 的完整性。在本报告中,我们评估了死后延迟、濒死状态和年龄对基因表达的影响,以及 pH 值和 RNA 完整性数量作为基因表达预测指标的效用,这些指标是在 1266 个 Affymetrix Exon Arrays 上测量的。我们使用 QuantiGene 评估了阵列数据的准确性,作为一种独立的非 PCR 方法。这些质量控制参数将允许数据库用户评估数据的准确性。我们报告说,在本研究的参数范围内,死后延迟、濒死状态和年龄对阵列质量的影响很小,阵列数据对可变的 RNA 完整性具有很强的鲁棒性,脑 pH 值对阵列性能的影响很小。QuantiGene 给出了与阵列数据非常相似的表达谱。这项研究是我们使人类、区域大脑表达免费获取的倡议的第一步。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac9c/3664422/95c31b0aee53/jnc0119-0275-f1.jpg

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