EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES
Currently there are several methods to extract bacterial DNA based on different principles. However, the amount and the quality of the DNA obtained by each one of those methods is highly variable and microorganism dependent, as illustrated by coagulase-negative staphylococci (CoNS) which have a thic...
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Universidade de São Paulo
2014-01-01
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doaj-fd101dd7726e473fb99335763b24b5542020-11-24T23:36:49ZengUniversidade de São PauloRevista do Instituto de Medicina Tropical de São Paulo1678-99462014-01-01561293310.1590/S0036-46652014000100004S0036-46652014000100029EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATESCaio Fernando de OliveiraThiago Galvao da Silva PaimKeli Cristine ReiterAlexandre RiegerPedro Alves D'azevedoCurrently there are several methods to extract bacterial DNA based on different principles. However, the amount and the quality of the DNA obtained by each one of those methods is highly variable and microorganism dependent, as illustrated by coagulase-negative staphylococci (CoNS) which have a thick cell wall that is difficult to lyse. This study was designed to compare the quality and the amount of CoNS DNA, extracted by four different techniques: two in-house protocols and two commercial kits. DNA amount and quality determination was performed through spectrophotometry. The extracted DNA was also analyzed using agarose gel electrophoresis and by PCR. 267 isolates of CoNS were used in this study. The column method and thermal lyses showed better results with regard to DNA quality (mean ratio of A260/280 = 1.95) and average concentration of DNA (), respectively. All four methods tested provided appropriate DNA for PCR amplification, but with different yields. DNA quality is important since it allows the application of a large number of molecular biology techniques, and also it's storage for a longer period of time. In this sense the extraction method based on an extraction column presented the best results for CoNS.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46652014000100029&lng=en&tlng=enDNA extractionCoagulase-negative Staphylococci (CoNS)Spectrophotometry |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Caio Fernando de Oliveira Thiago Galvao da Silva Paim Keli Cristine Reiter Alexandre Rieger Pedro Alves D'azevedo |
spellingShingle |
Caio Fernando de Oliveira Thiago Galvao da Silva Paim Keli Cristine Reiter Alexandre Rieger Pedro Alves D'azevedo EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES Revista do Instituto de Medicina Tropical de São Paulo DNA extraction Coagulase-negative Staphylococci (CoNS) Spectrophotometry |
author_facet |
Caio Fernando de Oliveira Thiago Galvao da Silva Paim Keli Cristine Reiter Alexandre Rieger Pedro Alves D'azevedo |
author_sort |
Caio Fernando de Oliveira |
title |
EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES |
title_short |
EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES |
title_full |
EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES |
title_fullStr |
EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES |
title_full_unstemmed |
EVALUATION OF FOUR DIFFERENT DNA EXTRACTION METHODS IN COAGULASE-NEGATIVE STAPHYLOCOCCI CLINICAL ISOLATES |
title_sort |
evaluation of four different dna extraction methods in coagulase-negative staphylococci clinical isolates |
publisher |
Universidade de São Paulo |
series |
Revista do Instituto de Medicina Tropical de São Paulo |
issn |
1678-9946 |
publishDate |
2014-01-01 |
description |
Currently there are several methods to extract bacterial DNA based on different principles. However, the amount and the quality of the DNA obtained by each one of those methods is highly variable and microorganism dependent, as illustrated by coagulase-negative staphylococci (CoNS) which have a thick cell wall that is difficult to lyse. This study was designed to compare the quality and the amount of CoNS DNA, extracted by four different techniques: two in-house protocols and two commercial kits. DNA amount and quality determination was performed through spectrophotometry. The extracted DNA was also analyzed using agarose gel electrophoresis and by PCR. 267 isolates of CoNS were used in this study. The column method and thermal lyses showed better results with regard to DNA quality (mean ratio of A260/280 = 1.95) and average concentration of DNA (), respectively. All four methods tested provided appropriate DNA for PCR amplification, but with different yields. DNA quality is important since it allows the application of a large number of molecular biology techniques, and also it's storage for a longer period of time. In this sense the extraction method based on an extraction column presented the best results for CoNS. |
topic |
DNA extraction Coagulase-negative Staphylococci (CoNS) Spectrophotometry |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46652014000100029&lng=en&tlng=en |
work_keys_str_mv |
AT caiofernandodeoliveira evaluationoffourdifferentdnaextractionmethodsincoagulasenegativestaphylococciclinicalisolates AT thiagogalvaodasilvapaim evaluationoffourdifferentdnaextractionmethodsincoagulasenegativestaphylococciclinicalisolates AT kelicristinereiter evaluationoffourdifferentdnaextractionmethodsincoagulasenegativestaphylococciclinicalisolates AT alexandrerieger evaluationoffourdifferentdnaextractionmethodsincoagulasenegativestaphylococciclinicalisolates AT pedroalvesdazevedo evaluationoffourdifferentdnaextractionmethodsincoagulasenegativestaphylococciclinicalisolates |
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