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一种用于测量RNA质量的新方法的评估。

Evaluation of a novel approach for the measurement of RNA quality.

作者信息

Wilkes Timothy M, Devonshire Alison S, Ellison Stephen Lr, Foy Carole A

机构信息

LGC, Queens Road, Teddington, Middlesex, TW11 0LY, UK.

出版信息

BMC Res Notes. 2010 Apr 1;3:89. doi: 10.1186/1756-0500-3-89.

Abstract

BACKGROUND

Microarray data interpretation can be affected by sample RNA integrity. The ScreenTape Degradation Value (SDV) is a novel RNA integrity metric specific to the ScreenTape(R) platform (Lab901). To characterise the performance of the ScreenTape(R) platform for RNA analysis and determine the robustness of the SDV metric, a panel of intentionally degraded RNA samples was prepared. These samples were used to evaluate the ScreenTape(R) platform against an alternative approach for measuring RNA integrity (Agilent Bioanalyzer RIN value). The samples were also subjected to microarray analysis and the resulting data correlated to the RNA integrity metrics.

FINDINGS

Measurement of SDV for a panel of intentionally degraded RNA samples ranged from 0 for intact RNA to 37 for degraded RNA, with corresponding RIN values ranging from 10 to 4 for the same set of samples. SDV and RIN scales both demonstrated comparable discrimination between differently treated samples (RIN 10 to 7, SDV 0 to 15), with the SDV exhibiting better discrimination at higher degradation levels. Increasing SDV values correlated with a decrease in microarray sample labelling efficiency and an increase in numbers of differentially expressed genes.

CONCLUSIONS

The ScreenTape(R) platform is comparable to the Bioanalyzer platform in terms of reproducibility and discrimination between different levels of RNA degradation. The robust nature of the SDV metric qualifies it as an alternative metric for RNA sample quality control, and a useful predictor of downstream microarray performance.

摘要

背景

微阵列数据的解读可能会受到样本RNA完整性的影响。ScreenTape降解值(SDV)是一种专门针对ScreenTape(R)平台(Lab901)的新型RNA完整性指标。为了表征ScreenTape(R)平台在RNA分析方面的性能,并确定SDV指标的稳健性,制备了一组故意降解的RNA样本。这些样本用于将ScreenTape(R)平台与另一种测量RNA完整性的方法(安捷伦生物分析仪RIN值)进行比较评估。这些样本还进行了微阵列分析,所得数据与RNA完整性指标相关。

研究结果

一组故意降解的RNA样本的SDV测量值范围从完整RNA的0到降解RNA的37,同一组样本的相应RIN值范围从10到4。SDV和RIN量表在不同处理样本之间均表现出可比的区分能力(RIN 10至7,SDV 0至15),其中SDV在较高降解水平下表现出更好的区分能力。SDV值的增加与微阵列样本标记效率的降低和差异表达基因数量的增加相关。

结论

ScreenTape(R)平台在可重复性和区分不同程度RNA降解方面与生物分析仪平台相当。SDV指标的稳健性使其有资格作为RNA样本质量控制的替代指标,以及下游微阵列性能的有用预测指标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfe3/2907863/6f50595b2b36/1756-0500-3-89-1.jpg

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