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PLUSTM Reagent

PLUS TM试剂

Company: Thermo Fisher Scientific
Catalog#: 11514015
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Stable Interference by shRNA with pSUPER.retro Vectors and Lipofectamine
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2012-08-05
[Abstract]  RNA interference is a powerful genetic approach for efficiently silencing target genes. Expression of short hairpin RNAs (shRNAs) allows analysis of the consequences of stably silencing genes. This protocol describes a method to stably integrate shRNA constructs with pSUPER.retro vectors and Lipofectamine in RKO and H1299 cells. This method can be applied to cells from other lines with modification of drug selection and cell conditions. pSUPER.retro vectors can be also transfected into packaging cells by this same method to produce retroviral supernatants. [摘要]  RNA干扰是用于有效沉默靶基因的强有力的遗传方法。 短发夹RNA(shRNA)的表达允许分析稳定沉默基因的后果。 此协议描述了稳定整合shRNA构建体与pSUPER.retro载体和Lipofectamine在RKO和H1299细胞中的方法。 这种方法可以应用于来自其他细胞系的药物选择和细胞条件的修改。 也可以通过相同的方法将pSUPER.retro载体转染到包装细胞中以产生逆转录病毒上清液。

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