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Journal of Biomolecular Screening
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1087057107312032v1
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Article

An Efficient and Fully Automated High-Throughput Transfection Method for Genome-Scale siRNA Screens

Namjin Chung*, Louis Locco, Kevin W. Huff, Steven Bartz, Peter S. Linsley, Marc Ferrer, and Berta Strulovici

Merck & Co.

* To whom correspondence should be addressed. E-mail: namjin.chung{at}gmail.com.


  Abstract
RNA interference (RNAi), combined with the availability of genome sequences, provides an unprecedented opportunity for the massive and parallel investigations of gene function. Small interfering RNA (siRNA) represents a popular and quick approach of RNAi for in vitro loss-of-function genetic screens. Efficient transfection of siRNA is critical for unambiguous interpretation of screen results and thus overall success of any siRNA screen. A high-throughput, lipid-based transfection method for siRNA was developed that can process eighty 384-well microplates in triplicate (for a total of 30,720 unique transfections) in 8 h. Transfection throughput was limited only by the speed of robotics, whereas the cost of screening was reduced. As a proof of principle, a genome-scale screen with a library of 22,108 siRNAs was performed to identify the genes sensitizing cells to mitomycin C at concentrations of 0, 20, and 60 nM. Transfection efficiency, performances of control siRNAs, and other quality metrics were monitored and demonstrated that the new, optimized transfection protocol produced high-quality results throughout the screen. (Journal of Biomolecular Screening XXXX:xx-xx)

First published on January 23, 2008, doi:10.1177/1087057107312032

Journal of Biomolecular Screening 2008;13:142.

A more recent version of this article appeared on February 1, 2008

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