Source:http://linkedlifedata.com/resource/pubmed/id/18216392
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
2008-3-4
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pubmed:abstractText |
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.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
1087-0571
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
13
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
142-8
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pubmed:dateRevised |
2011-5-23
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pubmed:meshHeading |
pubmed-meshheading:18216392-Algorithms,
pubmed-meshheading:18216392-Automation,
pubmed-meshheading:18216392-Efficiency,
pubmed-meshheading:18216392-Gene Expression Profiling,
pubmed-meshheading:18216392-Genome, Human,
pubmed-meshheading:18216392-HeLa Cells,
pubmed-meshheading:18216392-Humans,
pubmed-meshheading:18216392-RNA, Small Interfering,
pubmed-meshheading:18216392-Transfection
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pubmed:year |
2008
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pubmed:articleTitle |
An efficient and fully automated high-throughput transfection method for genome-scale siRNA screens.
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pubmed:affiliation |
Department of Automated Biotechnology, Merck Research Laboratories, North Wales, PA 19454, USA. namjin.chung@bms.com
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pubmed:publicationType |
Journal Article,
Evaluation Studies
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