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

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 descri...

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Bibliographic Details
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
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Summary: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.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-011-0693-6