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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2015.tde-31082015-140037
Document
Author
Full name
Maicon Ricardo Zieberg Passini
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Maldonado, Gabriel Padilla (President)
Sato, Maria Inês Zanoli
Vicente, Elisabete Jose
Title in Portuguese
Aperfeiçoamento de uma técnica para extrair DNA de água do mar e comparação de métodos de extração de DNA na caracterização de uma comunidade bacteriana marinha.
Keywords in Portuguese
DGGE
Ecologia molecular microbiana
Extração de DNA de água do mar
Micro-organismos marinhos
Abstract in Portuguese
Com as limitações para cultivar os micro-organismos, tornou-se necessário caracterizar molecularmente os recursos genéticos microbianos. Entretanto, como estes estudos são realizados através do DNA, podemos encontrar distintos resultados dependendo da metodologia de extração de DNA utilizada. Assim, essa pesquisa buscou aperfeiçoar uma técnica para extrair DNA de água do e para demonstrar a importância dos métodos de extração de DNA no estudo dos micro-organismos por métodos independentes de cultivo, foi caracterizada, através da técnica de DGGE, a diversidade de uma comunidade bacteriana marinha usando diferentes métodos de extração de DNA. O aperfeiçoamento da metodologia de extração de DNA de água do mar resultou em um protocolo com a qualidade do DNA similar à do protocolo original, com rendimento melhor e aproximadamente quatro vezes mais rápido. Em relação à caracterização, verificamos, pelo DGGE, distintos perfis da comunidade microbiana marinha, tanto em relação à presença e ausência das bandas de DNA, como em relação à sua intensidade.
Title in English
The improvement of a technique for seawater DNA extracting and the comparison of DNA extraction methods for the characterization of a marine bacterial community.
Keywords in English
DGGE
DNA extraction from seawater
Marine microorganisms
Molecular microbial ecology
Abstract in English
With the limitations of microorganisms culture techniques, it has become necessary to use molecular techniques to characterize microbial genetic resources. However, these studies are done using extracted environmental DNA, therefore we can find different results depending on the methodology chosen for the DNA extraction. Thus, this study aimed to improve a technique for extracting DNA from seawater and to demonstrate the importance of the DNA extraction methods in the study of microorganisms by cultivation-independent methods, it was characterized, by DGGE technique, the diversity of a marine bacterial community using different methods of DNA extraction. The improved methodology of DNA extraction from seawater resulted in a protocol similar in DNA quality, with better yield and approximately four times faster than the protocol initially described. Regarding the characterization by different DNA extraction protocols, we found distinct marine microbial community profiles, in relation the absence and presence of DNA bands and their relative intensity.
 
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Publishing Date
2015-09-04
 
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