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快速纯化兔粘液瘤病毒 DNA。

Rapid purification of myxoma virus DNA.

机构信息

Instituto Universitario de Biotecnología de Asturias, Departamento de Bioquímica y Biología Molecular, Edificio Santiago Gascón, Campus El Cristo, Universidad de Oviedo, 33006 Oviedo, Spain.

出版信息

J Virol Methods. 2009 Dec;162(1-2):284-7. doi: 10.1016/j.jviromet.2009.08.003. Epub 2009 Aug 20.

Abstract

A rapid and efficient procedure for the purification of myxoma virus DNA from infected cell cultures is described. The traditional method used for recovery of myxoma virus DNA involves multiple freeze-thawing cycles to disrupt cells and release virions followed by ultracentrifugation to concentrate virions for DNA extraction. Freeze-thaw cycles are time consuming and reduce viral titers, while ultracentrifugation steps limit the number of samples that can be processed at one time, reducing efficiency. In this report an optimized method circumventing the time-consuming techniques and replacing them with rapid, efficient steps adequate for the processing of larger numbers of samples is described. The traditional method was compared with the optimized protocol, which was found to be more efficient in terms of time required to complete the process and in the quantities of DNA purified.

摘要

本文描述了一种从感染细胞培养物中快速高效纯化兔粘液瘤病毒 DNA 的方法。传统的兔粘液瘤病毒 DNA 回收方法涉及多次冻融循环以破坏细胞并释放病毒粒子,然后进行超速离心以浓缩病毒粒子用于 DNA 提取。冻融循环耗时且降低病毒滴度,而超速离心步骤限制了一次可处理的样品数量,降低了效率。在本报告中,描述了一种优化的方法,该方法绕过耗时的技术,代之以快速、高效的步骤,足以处理更多数量的样品。传统方法与优化方案进行了比较,结果表明优化方案在完成整个过程所需的时间和纯化的 DNA 量方面更有效。

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