如何制备转染级质粒DNA

转染效率受各种不同参数的影响。�Besides factors such as cell culture, medium and vectors, one of the most critical parameters is DNA quality.� We prepared pEGF-N3 plasmid DNA with large plasmid preparation kits from three different vendors and tested transfection efficiencies by delivering pEGFP-N3 DNAs prepared by different methods to NIH-3T3 cells.�

我们使用Endofree Maxi plasmid Kit (Qiagen)、PerfectPrep Endofree plasmid Maxi Kit (5 PRIME, Inc.)和NucleoBond DNA Maxi试剂盒(machery - nagel GmbH & Co. KG。�The DNA quality was determined by measuring absorbance at 230, 260, 280 and 320 nm by spectrometry.
三种方法制备的DNA纯度如下:
MACHEREY-NAGEL>5、Prime > Qiagen

然后，pEGFP DNA通过PolyJet (Cat # SL100688) DNA转染试剂递送到NIH-3T3细胞，按照制造商的协议。根据DNA纯度结果，我们发现DNA产生于MACHEREY-NAGEL试剂盒给出的分数最高转染效率最低，而Qiagen的DNA制备试剂盒的效率最低。因此，质粒制备试剂盒从machery - nagel GmbH & Co. KG是制备转染级DNA的最佳选择。

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