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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. ...
[摘要] 用DNA转染细胞是分子生物学中一个不可缺少的方法。虽然脂转染试剂已实现商业化，此外还有一项十分快速，简便，高效，廉价，既通过磷酸钙与DNA的共沉淀转染真核细胞的方法。（1）DNA附着在不可溶的磷酸钙沉淀上，粘合到细胞表面后，共同通过内吞作用被带入细胞内部。经过优化的磷酸钙转染被广泛应用用许多粘合或不粘合的细胞系中。（2）DNA进入靶细胞，可以是瞬时的也可以建立稳定的细胞系，这个方法被广泛用于组装病毒。就不同的细胞株转染效率可以接近100％。这里GFP的质粒和HEK293细胞为例，介绍利用CaPO4转染的具体方法。
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