Johny Tina Kollannoor, Bhat Sarita Ganapathy
Department of Biotechnology, Cochin University of Science and Technology, Kalamassery, Cochin, 682022, Kerala, India.
Methods Mol Biol. 2017;1620:249-265. doi: 10.1007/978-1-4939-7060-5_17.
Metagenomics approach involves direct genetic analysis of environmental samples, evading the tedious culturing process. Polymerase chain reaction is one invaluable tool used for such analyses. Here, we describe one protocol for metagenomic DNA isolation that gives inhibitor-free DNA suitable for PCR and other genetic manipulations. Subsequently, the chapter describes the use of PCR as an indicator of quality of DNA and to amplify a marker of phylogeny. Further, the application of PCR for detection of specific genes and screening of metagenomic libraries is outlined.
宏基因组学方法涉及对环境样本进行直接基因分析,避免了繁琐的培养过程。聚合酶链反应是用于此类分析的一种非常有价值的工具。在这里,我们描述了一种宏基因组DNA分离方案,该方案可提供适用于PCR和其他基因操作的无抑制剂DNA。随后,本章介绍了使用PCR作为DNA质量指标并扩增系统发育标记的方法。此外,还概述了PCR在检测特定基因和筛选宏基因组文库中的应用。