Department de Biologia Celular, Instituto Central de Ciências Sul, Universidade de Brasília (UnB), Brasília, DF, Brazil.
Biotechnol Lett. 2011 Nov;33(11):2169-75. doi: 10.1007/s10529-011-0693-6. Epub 2011 Jul 12.
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.
元基因组研究的一个挑战在于从环境样本中提取和纯化 DNA。巴西塞拉多地区的土壤在 DNA 提取方面存在一些技术难题：高粘土含量（>55%w/w）、低 pH 值（4.7）和高铁含量（146ppm）。在这里，我们首次描述了从这些特殊土壤特性中有效回收和纯化与微生物 DNA 相关的方法，并构建和验证了两个元基因组文库：一个约 8kb 插入大小的 150,000 个克隆文库和一个约 35kb 插入大小的 65,000 个克隆文库。这些元基因组文库的构建将允许对来自这一濒危生物群落的土壤中存在的微生物群落进行生物技术开发。
