RRC ID |
6830
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著者 |
Fujita S, Takano K, Ota E, Sano T, Yoshikawa T, Miyake M, Miyake J.
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タイトル |
New methods for reverse transfection with siRNA from a solid surface.
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ジャーナル |
Methods Mol Biol
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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
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リソース情報 |
ヒト・動物細胞 |
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