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使用2100生物分析仪毛细管电泳系统通过测量RNA完整性来预测寡核苷酸微阵列杂交的定性结果。

Prediction of qualitative outcome of oligonucleotide microarray hybridization by measurement of RNA integrity using the 2100 Bioanalyzer capillary electrophoresis system.

作者信息

Kiewe Philipp, Gueller Saskia, Komor Martina, Stroux Andrea, Thiel Eckhard, Hofmann Wolf-Karsten

机构信息

Department of Hematology, Oncology, and Transfusion Medicine, Charité-University Hospital Benjamin Franklin, Hindenburgdamm 30, 12203 Berlin, Germany.

出版信息

Ann Hematol. 2009 Dec;88(12):1177-83. doi: 10.1007/s00277-009-0751-5. Epub 2009 May 8.

Abstract

RNA quality is critical to achieve valid results in microarray experiments and to save resources. The RNA integrity number (RIN) can be measured with minimal sample consumption by microfluidics-based capillary electrophoresis. To determine whether RIN can predict the qualitative outcome of microarray hybridization, we measured RIN in total RNA samples from 484 different experiments by the 2100 Bioanalyzer system and correlated with the percentage of present calls (%pc) of downstream oligonucleotide microarrays. The correlation coefficient for RNA and %pc in all 408 samples for which the bioanalyzer algorithm was able to produce an RIN was 0.475 (p < 0.05), ranging from 0.039 to 0.673 for different tissue- and assay-type subgroups. Multivariate analysis found RIN to be the best predictor of microarray quality as assessed by %pc, outperforming the 28S to 18S ratio. For a %pc threshold of 25% and 35%, we determined optimal cut points for RIN at 7.15 and 8.05, respectively. Using the suggested cut points, RIN can support the final decision whether a certain RNA sample is appropriate for successful microarray hybridization.

摘要

RNA质量对于在微阵列实验中获得有效结果并节省资源至关重要。RNA完整性数值(RIN)可以通过基于微流控的毛细管电泳以最少的样本消耗进行测量。为了确定RIN是否能够预测微阵列杂交的定性结果,我们使用2100生物分析仪系统测量了来自484个不同实验的总RNA样本中的RIN,并将其与下游寡核苷酸微阵列的检出率(%pc)进行关联。对于生物分析仪算法能够得出RIN的所有408个样本,RNA与%pc的相关系数为0.475(p < 0.05),不同组织和检测类型亚组的相关系数范围为0.039至0.673。多变量分析发现,以%pc评估时,RIN是微阵列质量的最佳预测指标,优于28S与18S的比率。对于25%和35%的%pc阈值,我们分别确定RIN的最佳切点为7.15和8.05。使用建议的切点,RIN可以辅助做出关于某个RNA样本是否适合成功进行微阵列杂交的最终决定。

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