Armstrong B, Garner H R
Institute for Development of Advanced Technology, General Atomics, San Diego, CA 92186.
Genet Anal Tech Appl. 1992 Oct-Dec;9(5-6):127-33. doi: 10.1016/1050-3862(92)90036-5.
A modified alkaline lysis method for preparing plasmid DNA from bacterial cells has been developed for automation implementation. The objective of this study was to develop a simplified centrifuge-based protocol that can process samples in a microwell plate. These manual experiments and parametric studies show that the alkaline lysis method can be modified significantly to enable DNA preparations to be done rapidly and reliably by an automated system. The conclusions of this study were (a) centrifugation at < 1500 g is sufficient, (b) centrifugation times need not be extended to compensate for the reduced force, (c) reactions can be done at room temperature, (d) reagent volumes can be reduced over those typically used, and (e) certain reagents can be combined to simplify the handling of fluids.
一种用于从细菌细胞中制备质粒DNA的改良碱性裂解方法已被开发出来用于自动化实施。本研究的目的是开发一种基于离心机的简化方案，该方案可在微孔板中处理样品。这些手动实验和参数研究表明，碱性裂解方法可以进行显著修改，以使DNA制备能够通过自动化系统快速且可靠地完成。本研究的结论是：（a）在<1500 g下离心就足够了；（b）不需要延长离心时间来补偿降低的离心力；（c）反应可以在室温下进行；（d）试剂体积可以比通常使用的体积减少；（e）某些试剂可以合并以简化液体处理。
