Shamim Kashif, Sharma Jaya, Dubey Santosh Kumar
Laboratory of Bacterial Genetics and Environmental Biotechnology, Department of Microbiology, Goa University, Taleigao Plateau, Goa, 403206, India.
3 Biotech. 2017 Jul;7(3):182. doi: 10.1007/s13205-017-0846-y. Epub 2017 Jun 29.
Metagenomic DNA from sediments of selective estuaries of Goa, India was extracted using a simple, fast, efficient and environment friendly method. The recovery of pure metagenomic DNA from our method was significantly high as compared to other well-known methods since the concentration of recovered metagenomic DNA ranged from 1185.1 to 4579.7 µg/g of sediment. The purity of metagenomic DNA was also considerably high as the ratio of absorbance at 260 and 280 nm ranged from 1.88 to 1.94. Therefore, the recovered metagenomic DNA was directly used to perform various molecular biology experiments viz. restriction digestion, PCR amplification, cloning and metagenomic library construction. This clearly proved that our protocol for metagenomic DNA extraction using silica gel efficiently removed the contaminants and prevented shearing of the metagenomic DNA. Thus, this modified method can be used to recover pure metagenomic DNA from various estuarine sediments in a rapid, efficient and eco-friendly manner.
采用一种简单、快速、高效且环境友好的方法，从印度果阿邦特定河口的沉积物中提取了宏基因组DNA。与其他知名方法相比，我们的方法所回收的纯宏基因组DNA产量显著更高，因为回收的宏基因组DNA浓度范围为每克沉积物1185.1至4579.7微克。宏基因组DNA的纯度也相当高，260和280纳米处吸光度的比值在1.88至1.94之间。因此，回收的宏基因组DNA可直接用于进行各种分子生物学实验，即限制性消化、PCR扩增、克隆和宏基因组文库构建。这清楚地证明，我们使用硅胶提取宏基因组DNA的方案能有效去除污染物，并防止宏基因组DNA被剪切。因此，这种改良方法可用于以快速、高效且环保的方式从各种河口沉积物中回收纯宏基因组DNA。
