Improved method for the extraction of high-quality DNA from lignocellulosic compost samples for metagenomic studies

被引:11
|
作者
Costa, Angela M. A. [1 ]
Santos, Andreia O. [1 ]
Sousa, Joana [1 ]
Rodrigues, Joana L. [1 ]
Gudina, Eduardo J. [1 ]
Silverio, Sara C. [1 ]
Rodrigues, Ligia R. [1 ]
机构
[1] Univ Minho, CEB Ctr Biol Engn, P-4710057 Braga, Portugal
关键词
Lignocellulosic; Compost; Metagenomic DNA; Humic acids; Activated charcoal; PURIFICATION PROCEDURES; ENVIRONMENTAL DNA; HUMIC SUBSTANCES; ORGANIC WASTES; SOIL; OPTIMIZATION; DISCOVERY; EFFICIENT; RECOVERY; BIOMASS;
D O I
10.1007/s00253-021-11647-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The world economy is currently moving towards more sustainable approaches. Lignocellulosic biomass has been widely used as a substitute for fossil sources since it is considered a low-cost bio-renewable resource due to its abundance and continuous production. Compost habitats presenting high content of lignocellulosic biomass are considered a promising source of robust lignocellulose-degrading enzymes. Recently, several novel biocatalysts from different environments have been identified using metagenomic techniques. A key point of the metagenomics studies is the extraction and purification of nucleic acids. Nevertheless, the isolation of high molecular weight DNA from soil-like samples, such as compost, with the required quality for metagenomic approaches remains technically challenging, mainly due to the complex composition of the samples and the presence of contaminants like humic substances. In this work, a rapid and cost-effective protocol for metagenomic DNA extraction from compost samples composed of lignocellulosic residues and containing high content of humic substances was developed. The metagenomic DNA was considered as representative of the global environment and presented high quality (> 99% of humic acids effectively removed) and sufficient quantity (10.5-13.8 mu g g(-1) of compost) for downstream applications, namely functional metagenomic studies. The protocol takes about 4 h of bench work, and it can be performed using standard molecular biology equipment and reagents available in the laboratory.
引用
收藏
页码:8881 / 8893
页数:13
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