title: Comparison of DNA extraction protocols for microbial communities from soil treated with biochar.
abstract: Many studies have evaluated the effects of biochar application on soil structure and plant growth. However, there are very few studies describing the effect of biochar on native soil microbial communities. Microbial analysis of environmental samples requires accurate and reproducible methods for the extraction of DNA from samples. Because of the variety among microbial species and the strong adsorption of the phosphate backbone of the DNA molecule to biochar, extracting and purifying high quality microbial DNA from biochar-amended soil is not a trivial process and can be considerably more difficult than the extraction of DNA from other environmental samples. The aim of this study was to compare the relative efficacies of three commercial DNA extraction kits, the FastDNA® SPIN Kit for Soil (FD kit), the PowerSoil® DNA Isolation Kit (PS kit) and the ZR Soil Microbe DNA Kit Miniprep™ (ZR kit), for extracting microbial genomic DNA from sand treated with different types of biochar. The methods were evaluated by comparing the DNA yields and purity and by analysing the bacterial and fungal community profiles generated by PCR-DGGE. Our results showed that the PCR-DGGE profiles for bacterial and fungal communities were highly affected by the purity and yield of the different DNA extracts. Among the tested kits, the PS kit was the most efficient with respect to the amount and purity of recovered DNA and considering the complexity of the generated DGGE microbial fingerprint from the sand-biochar samples.
authors.affiliation: Laboratório de Ecologia Microbiana Molecular Instituto de Microbiologia Prof. Paulo de Góes Universidade Federal do Rio de Janeiro Rio de JaneiroRJ Brazil.
authors.forename: D C A
authors.initials: DC
authors.lastname: Leite
authors.affiliation: Empresa Brasileira de Pesquisa Agropecuária Solos Rio de JaneiroRJ Brazil.
authors.forename: F C
authors.initials: FC
authors.lastname: Balieiro
authors.affiliation: Empresa Brasileira de Pesquisa Agropecuária Solos Rio de JaneiroRJ Brazil.
authors.forename: C A
authors.initials: CA
authors.lastname: Pires
authors.affiliation: Empresa Brasileira de Pesquisa Agropecuária Arroz e Feijão GoiásGO Brazil.
authors.forename: B E
authors.initials: BE
authors.lastname: Madari
authors.affiliation: Laboratório de Ecologia Microbiana Molecular Instituto de Microbiologia Prof. Paulo de Góes Universidade Federal do Rio de Janeiro Rio de JaneiroRJ Brazil.
authors.forename: A S
authors.initials: AS
authors.lastname: Rosado
authors.affiliation: Empresa Brasileira de Pesquisa Agropecuária Solos Rio de JaneiroRJ Brazil.
authors.forename: H L C
authors.initials: HL
authors.lastname: Coutinho
authors.affiliation: Laboratório de Ecologia Microbiana Molecular Instituto de Microbiologia Prof. Paulo de Góes Universidade Federal do Rio de Janeiro Rio de JaneiroRJ Brazil.
authors.forename: R S
authors.initials: RS
authors.lastname: Peixoto
mesh_terms: D058448:Biota; D002606:Charcoal; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D058645:Denaturing Gradient Gel Electrophoresis; D016133:Polymerase Chain Reaction; D012987:Soil / Q000737:chemistry*; D012988:Soil Microbiology
chemical_list: D004269:DNA, Bacterial; D004271:DNA, Fungal; D012987:Soil; C540010:biochar; D002606:Charcoal
keywords: DNA extraction; DNA purity indices; PCR-DGGE; biochar; microbial communities