Smith G J, Helf M, Nesbet C, Betita H A, Meek J, Ferre F
Althea Technologies, San Diego, CA, USA.
Biotechniques. 1999 Mar;26(3):518-22, 524, 526. doi: 10.2144/99263rr03.
Plasmid DNA is being used successfully as a gene delivery vector in a variety of clinical applications. Similar to other pharmaceutical products for clinical use, the plasmid vectors must meet rigorous purity standards. One important contaminant is the DNA of the host cell used to produce the plasmids. We have developed a new method to accurately quantitate E. coli host-cell DNA in plasmid preparations. This method is based on kinetic PCR using the ABI PRISM 7700 with 23S rDNA as a target. This precise assay is significantly faster and has a lower limit of quantitation than the currently used Southern-based methods.
质粒DNA作为一种基因传递载体已成功应用于多种临床领域。与其他临床用药品类似，质粒载体必须符合严格的纯度标准。一个重要的污染物是用于生产质粒的宿主细胞的DNA。我们开发了一种新方法，可准确测定质粒制剂中大肠杆菌宿主细胞DNA的含量。该方法基于使用ABI PRISM 7700以23S rDNA为靶标的动力学PCR。与目前使用的基于Southern杂交的方法相比，这种精确的检测方法速度明显更快，定量下限更低。
