Construction and validation of two metagenomic DNA libraries from Cerrado soil with high clay content

• Published in: Biotechnology letters 2011-11, Vol.33 (11), p.2169-2175
• Main Authors: de Castro, Alinne Pereira, Quirino, Betania Ferraz, Allen, Heather, Williamson, Lynn L, Handelsman, Jo, Krüger, Ricardo Henrique
• Format: Article
• Language: English
• Subjects: Aluminum Silicates; Applied Microbiology; Biochemistry; Biodiversity; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Brazil; cerrado soils; Clay; clones; Construction; Deoxyribonucleic acid; DNA; ecosystems; Extraction; Fundamental and applied biological sciences. Psychology; Gene Library; genomic libraries; Genomics; Hydrogen-Ion Concentration; Inserts; Iron - analysis; Libraries; Life Sciences; Metagenome; metagenomics; microbial communities; Microbiology; Microorganisms; Original Research Paper; Purification; Soil (material); Soil - chemistry; Soil Microbiology; Soils
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-011-0693-6

A challenge of metagenomic studies is in the extraction and purification of DNA from environmental samples. The soils of the Cerrado region of Brazil present several technical difficulties to DNA extraction: high clay content (>55% w/w), low pH (4.7) and high iron levels (146 ppm). Here we describe for the first time the efficient recovery and purification of microbial DNA associated with these unusual soil characteristics and the construction and validation of two metagenomic libraries: a 150,000 clones library with insert size of approximately 8 kb and a 65,000 clones library with insert size of approximately 35 kb. The construction of these metagenomic libraries will allow the biotechnological exploitation of the microbial community present in the soil from this endangered biome.