GeneClip™ U1 Hairpin Cloning System－Puromycin

GeneClipTM U1 Hairpin Cloning Systems(a-f) (GeneClipTM U1发夹克隆系统)由线性质粒组成，这些质粒是为了在人类细胞中快速、简便地克隆人类目的序列，来表达短发夹RNAs(shRNAs)而设计的。转染至人类细胞中后，从包含U1 RNA聚合酶启动子和siRNA模板的DNA质粒可有效地进行短干扰RNAs(siRNAs)的体内表达。U1启动子通常用于体外成功地产生发夹siRNAs(1,2)。为使发夹siRNA插入pGeneClipTM 载体，将2个短DNA寡核苷酸经退火，形成包含该发夹siRNA的目的序列的DNA插入物。退火后，寡核苷酸形成与pGeneClipTM Vector末端匹配的突出末端，便于粘性末端连接。一经转染，RNA聚合酶II即在体内转录发夹插入序列，生成发夹siRNAs。

siRNA Target Designer 程序(www.promega.com/siRNADesigner/)可用来为使用Promega RNA干扰系统设计寡核苷酸序列。该程序对输入的DNA或RNA序列设计符合siRNA要求的区域进行分析。能够沉默这些序列的siRNAs将会与选定的试剂盒所要求的寡核苷酸序列一同显示出来。

特点

  •  更多载体选择: 这些系统提供的载体含有各种用于稳定转染真核细胞的抗生素抗性选择标记物，或用于确定转染效率的hMGFP。

  •  节省时间: 载体以预消化的形式提供以减少载体制备的时间。

  •  便利: 每个系统都包含T4 DNA 连接酶, 2X 快速连接缓冲液, 寡核苷酸退火缓冲液和pGeneClipTM载体。

  •  轻松鉴定目的克隆: 使用PstI 酶切快速鉴定阳性重组体。

应用

  •  人类细胞中的DNA介导的RNA干扰。

操作手册

Technical Manual      TM256

储存条件: 储存于–20°C.

参考文献

1. Novarino,G.et al.(2004)J.Neuroscience24,5322-30.

2. Denti,M-A.et al.(2004)Mol.Ther.10,191-9.

技术资料

RNA　Interference Systems Brochure   BR142

Promega Notes                        PN089

(a) For nonprofit and noncommercial research use only. With respect to commercial researchuse or use by for-profit organizations, or any diagnostic or therapeutic uses, please contactPromega for supply and licensing information. Please refer to the Acknowledgments/Disclaimers section in the Technical and Legal References chapter for complete terms.

(b) This product is covered under license from Carnegie Institution of Washington under U.S.Pat. No. 6,506,559, Australian Pat. No. 743798 and other patents pending. Commercial useof this product may require a separate license from Carnegie.

(c) Cat.# C8790: The CMV promoter and its use are covered under U.S. Pat. Nos. 5,168,062and 5,385,839 owned by the University of Iowa Research Foundation, Iowa City, Iowa, andlicensed FOR RESEARCH USE ONLY. Research Use includes contract research for whichmonetary or other consideration may be received. Other commercial users must obtain alicense to these patents directly from the University of Iowa Research Foundation