Department of Microbiology, Immunology and Molecular Genetics, University of California Los Angeles, Los Angeles, California, USA.
PLoS One. 2009 Nov 18;4(11):e7750. doi: 10.1371/journal.pone.0007750.
Purity, yield, speed and cost are important considerations in plasmid purification, but it is difficult to achieve all of these at the same time. Currently, there are many protocols and kits for DNA purification, however none maximize all four considerations.
METHODOLOGY/PRINCIPAL FINDINGS: We now describe a fast, efficient and economic in-house protocol for plasmid preparation using glass syringe filters. Plasmid yield and quality as determined by enzyme digestion and transfection efficiency were equivalent to the expensive commercial kits. Importantly, the time required for purification was much less than that required using a commercial kit.
CONCLUSIONS/SIGNIFICANCE: This method provides DNA yield and quality similar to that obtained with commercial kits, but is more rapid and less costly.
在质粒纯化中,纯度、产率、速度和成本都是重要的考虑因素,但很难同时实现所有这些因素。目前,有许多用于 DNA 纯化的方案和试剂盒,但没有一个方案能够最大化这四个方面的考虑。
方法/主要发现:我们现在描述了一种使用玻璃注射器过滤器快速、高效和经济的质粒制备的内部方案。通过酶消化和转染效率确定的质粒产量和质量与昂贵的商业试剂盒相当。重要的是,纯化所需的时间比使用商业试剂盒所需的时间要少得多。
结论/意义:这种方法提供了与商业试剂盒相似的 DNA 产量和质量,但速度更快,成本更低。