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Author:
Date:
2012-02-05
[Abstract] Transfection of DNA into cells is an indispensible protocol in molecular biology. While plenty of lipid-based transfection reagents are commercially available nowadays, a quick, simple, efficient and inexpensive method is to transfect eukaryotic cells via calcium phosphate co-precipitation with DNA (Graham and van der Eb, 1973). The insoluble calcium phosphate precipitate with the attached DNA adheres to the cell surface and is brought into the cells by endocytosis. Calcium phosphate transfection has been optimized and widely used with many adherent and nonadherent cell lines (Jordan et al., 1996). Calcium phosphate transfection can result in transient expression of the delivered DNA in the target cell, or establishment of stable cell lines (the latter requires a drug selection ...
[摘要] 小干扰RNA（siRNA）是一类20-25个核苷酸的双链RNA，在许多生物过程中起重要作用（Hamilton和Baulcombe，1999）。 siRNA通过"中和"靶蛋白的mRNA起作用，促进mRNA的降解，从而改变蛋白的生物效应（综述于Hannon和Rossi，2004）。 siRNA也可以改变调节RNA的细胞内水平。 在生物研究中使用siRNA操纵目的基因的表达通常被称为RNA干扰或敲除技术（Elbashir等人，2001）。 合成的siRNA是一种新兴的工具，现在广泛应用于这些研究。 不同公司采用多种算法来设计siRNA产品，其在功效，特异性和成本等方面不同。 在此使用来自Dharmacon的siGENOME SMARTpool试剂解释siRNA敲低的示例方案。
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