Abstract
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.
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This work was supported by grants from FAPDF and CNPq.
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10529_2011_693_MOESM1_ESM.eps
Supplementary Table 1 Classification of end sequenced clones from the low copy number plasmid and fosmid metagenomic library. Plasmid and fosmid DNA were isolated by using QIAprep Miniprep (QIagen) and sequenced with forward pCF430 primer and reverse pCF430 primer to low copy number plasmid. The fosmid ends were sequenced with PCC1FOS forward primer and PCC1FOS reverse primer. The high quality sequences of both libraries were submitted to search for homology in public databases using BLASTx. (EPS 2073 kb)
10529_2011_693_MOESM2_ESM.eps
Supplementary Fig. 1. Map of Brazil showing the localization of the Cerrado biome and how it has been reduced. a Original area covered with Cerrado vegetation and b Current area covered with Cerrado vegetation. Map of Brazil-Conservation International 2008 (http://www.conservation.org.br/). (EPS 693 kb)
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de Castro, A.P., Quirino, B.F., Allen, H. et al. Construction and validation of two metagenomic DNA libraries from Cerrado soil with high clay content. Biotechnol Lett 33, 2169–2175 (2011). https://doi.org/10.1007/s10529-011-0693-6
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DOI: https://doi.org/10.1007/s10529-011-0693-6