RRC ID 6830
著者 Fujita S, Takano K, Ota E, Sano T, Yoshikawa T, Miyake M, Miyake J.
タイトル New methods for reverse transfection with siRNA from a solid surface.
ジャーナル Methods Mol Biol
Abstract We describe two efficient and inexpensive methods for reverse transfection with siRNA from a solid surface. One method involves localized reverse transfection from spots on a glass slide, which is mainly useful for making "transfection microarrays" (TMAs). The other involves reverse transfection in multiple wells of microtiter plates. Conditions for cell culture, preparation of reagents, and details of reverse transfection have been determined for several lines of cells, but we focus here on experiments with HeLa cells. In particular, we evaluated the efficiency of transfection, the cytotoxic effects of reverse transfection, and the efficiency of gene "knockdown" by transfection. We also performed phenotypic screening for a functional gene, during which cell viability was evaluated in terms of fluorescence from Calcein-AM. Our methods for reverse transfection with siRNA should be powerful tools that are useful for high-throughput analysis of functional genes.
巻・号 623
ページ 197-209
公開日 2010-1-1
DOI 10.1007/978-1-60761-588-0_13
PMID 20217553
MeSH Gene Knockdown Techniques HeLa Cells Humans RNA, Small Interfering / genetics Transfection / methods*
引用数 4
WOS 分野 BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞