一种用磷酸钙法高效转染HEK293T细胞方法的建立Establishment of a High-Efficiency Calcium Phosphate Transfection Method for HEK 293T Cells
陈晓,蒋捍东,路新枝,于文功
摘要(Abstract):
利用磷酸钙转染法将lacZ-pShuttle质粒导入HEK 293T细胞中,实验证明,pH值、DNA纯度、转染后培养时间和甘油休克时间是影响转染效率的重要因素。转染后培养6h进行甘油休克效果最佳,比培养0h直接进行休克转染效率升高584.4%;甘油休克时间为4.5min时转染效果最佳,比不用甘油休克转染效率升高130.8%。通过对影响转染的几个条件进行优化,建立了1种可以高效转染HEK 293T细胞的方法,且简便易行。
关键词(KeyWords): 磷酸钙转染;甘油休克;X-gal染色
基金项目(Foundation): 山东省自然科学基金项目(Y2000C09)资助
作者(Author): 陈晓,蒋捍东,路新枝,于文功
DOI: 10.16441/j.cnki.hdxb.2005.05.020
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