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

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• 作者：Alinne Pereira de Castro (1) p.alinne@gmail.com
  Betania Ferraz Quirino (23) betaniaf@pos.ucb.br
  Heather Allen (4) heather.Allen@ARS.USDA.GOV
  Lynn L. Williamson (4) lynnlwilliamson@msn.com
  Jo Handelsman (45) jo.handelsman@yale.edu
  Ricardo Henrique Krüger (1) kruger@unb.br
• 关键词：Clay – DNA extraction – Iron – Library – Metagenomics – Soil
• 刊名：Biotechnology Letters
• 出版年：2011
• 出版时间：November 2011
• 年：2011
• 卷：33
• 期：11
• 页码：2169-2175
• 全文大小：1.7 MB
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• 作者单位：1. Laboratório de Enzimologia, Department de Biologia Celular, Instituto Central de Ciências Sul, Universidade de Brasília (UnB), Brasília, DF CEP 700910-900, Brazil2. Genomic Sciences and Biotechnology, Universidade Católica de Brasília, Brasília, DF, Brazil3. Embrapa-Agrienergy, Parque Esta??o Biológica, Brasília, DF 70770-901, Brazil4. Department of Plant Pathology, University of Wisconsin, 1630 Linden Dr. Madison, Madison, WI 53706, USA5. Department of Molecular, Cellular & Developmental Biology, Yale University, New Haven, CT 06511, USA
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

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.