Aranishi Futoshi
Molecular Biology Division, National Institute of Fisheries Science, Kanazawa, Yokohama, Japan.
Mol Biotechnol. 2002 May;21(1):39-41. doi: 10.1385/MB:21:1:039.
A scale-flexible and cost-efective protocol for plasmid preparation is described to cover miniprep and midiprep scale work in a microcentriguge format for analysis of recombinant clones, this protocol relies on a modified alkaline lysis of Escherichia coli cells and subsequent purification of plasmid DNA with no organic extraction and alcohol precipitation. It can process up to 20 mL of E. coli cells carrying 3-10 kbp plasmid vectors in < 10 min. Flexprep delivers sufficient yield and purity of plasmid DNA for routine applications including restriction enzyme digestion and fluorescent automated sequencing.
本文描述了一种适用于多种规模且经济高效的质粒制备方案，可涵盖微量制备和中量制备规模的工作，采用微量离心机形式用于重组克隆分析。该方案依赖于对大肠杆菌细胞进行改良的碱裂解，随后在不进行有机抽提和乙醇沉淀的情况下纯化质粒DNA。它能在不到10分钟的时间内处理多达20 mL携带3 - 10 kbp质粒载体的大肠杆菌细胞。Flexprep质粒制备方案能提供足够产量和纯度的质粒DNA，可用于包括限制性酶切消化和荧光自动测序在内的常规应用。
