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Novel Analytic Criteria and Effective Plate Designs for Quality Control in Genome-Scale RNAi Screens

Biometrics Research, Merck Research Laboratories, West Point, Pennsylvania, xiaohua_zhang{at}merck.com

One of the most fundamental challenges in genome-wide RNA interference (RNAi) screens is to glean biological significance from mounds of data, which relies on the development and adoption of appropriate analytic methods and designs for quality control (QC) and hit selection. Currently, a Z-factor-based QC criterion is widely used to evaluate data quality. However, this criterion cannot take into account the fact that different positive controls may have different effect sizes and leads to inconsistent QC results in experiments with 2 or more positive controls with different effect sizes. In this study, based on a recently proposed parameter, strictly standardized mean difference (SSMD), novel QC criteria are constructed for evaluating data quality in genome-wide RNAi screens. Two good features of these novel criteria are: (1) SSMD has both clear original and probability meanings for evaluating the differentiation between positive and negative controls and hence the SSMD-based QC criteria have a solid probabilistic and statistical basis, and (2) these QC criteria obtain consistent QC results for multiple positive controls with different effect sizes. In addition, I propose multiple plate designs and the guidelines for using them in genome-wide RNAi screens. Finally, I provide strategies for using the SSMD-based QC criteria and effective plate design together to improve data quality. The novel SSMD-based QC criteria, effective plate designs, and related guidelines and strategies may greatly help to obtain high quality of data in genome-wide RNAi screens. (Journal of Biomolecular Screening 2008:363-377)

Key Words: strictly standardized mean difference • Z factor • quality control • plate design • RNAi high-throughput screening

Journal of Biomolecular Screening, Vol. 13, No. 5, 363-377 (2008)
DOI: 10.1177/1087057108317062


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