Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis
Background and objectives: Given the difficulty in establishing an ideal method that can successfully extract bacterial deoxyribonucleic acid (DNA) in Gram-negative bacteria, the objective of this work was to compare protocols for the extraction of bacterial DNA and to report the more practical, fas...
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| Format: | Article |
| Language: | English |
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College of Medicine, Al-Nahrain University
2022-06-01
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| Series: | Baghdad Journal of Biochemistry and Applied Biological Sciences |
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| Online Access: | https://bjbabs.org/index.php/bjbabs/article/view/123 |
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| author | Oslaene Alves de Brito Francisca Alves dos Santos Tassia Thais Al Yafawi Cícero Roberto Nascimento Saraiva Rakel Olinda Macedo da Silva Lívia Maria Garcia Leandro Pedro Everson Alexandre de Aquino Dárcio Luiz de Sousa Júnior Maria Karollyna do Nascimento Silva Leandro |
| author_facet | Oslaene Alves de Brito Francisca Alves dos Santos Tassia Thais Al Yafawi Cícero Roberto Nascimento Saraiva Rakel Olinda Macedo da Silva Lívia Maria Garcia Leandro Pedro Everson Alexandre de Aquino Dárcio Luiz de Sousa Júnior Maria Karollyna do Nascimento Silva Leandro |
| author_sort | Oslaene Alves de Brito |
| collection | DOAJ |
| description | Background and objectives: Given the difficulty in establishing an ideal method that can successfully extract bacterial deoxyribonucleic acid (DNA) in Gram-negative bacteria, the objective of this work was to compare protocols for the extraction of bacterial DNA and to report the more practical, faster, and less costly ones.
Methods: Bacterial species used were Escherichia coli, provided by Dr. Leão Sampaio University Center. The methods used to extract bacterial DNA were: sodium dodecyl sulfate (SDS), salting-out, cetyltrimethylammonium bromide (CTAB)/Phenol-Chloroform, and commercial kit for DNA isolation from Promega. The extracted DNA by all the methods was detected and assessed by the agarose gel electrophoresis.
Results: The best result was that obtained by the SDS method, it showed the greatest advantage for bringing speed, low cost, and good concentration of the extracted material. The other methods showed low-quality DNA, probably due to the presence of large amounts of proteins in the cell wall of E. coli interfering with the quality of the samples.
Conclusions: It was concluded that the SDS method can provide better DNA in terms of quality, which is preferred for the amplification by the polymerase (PCR) chain reaction. This will be helpful for diagnostic microbiologists because of the low cost and good quality of the extracted DNA. |
| format | Article |
| id | doaj-art-ef7e8b3a15aa4963a15dab4e9b8de874 |
| institution | Kabale University |
| issn | 2706-9915 |
| language | English |
| publishDate | 2022-06-01 |
| publisher | College of Medicine, Al-Nahrain University |
| record_format | Article |
| series | Baghdad Journal of Biochemistry and Applied Biological Sciences |
| spelling | doaj-art-ef7e8b3a15aa4963a15dab4e9b8de8742025-08-20T03:47:23ZengCollege of Medicine, Al-Nahrain UniversityBaghdad Journal of Biochemistry and Applied Biological Sciences2706-99152022-06-013213314410.47419/bjbabs.v3i02.123Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresisOslaene Alves de Brito0Francisca Alves dos Santos1Tassia Thais Al Yafawi2Cícero Roberto Nascimento Saraiva3Rakel Olinda Macedo da Silva4Lívia Maria Garcia Leandro5Pedro Everson Alexandre de Aquino6Dárcio Luiz de Sousa Júnior7Maria Karollyna do Nascimento Silva Leandro8Doctor Leão Sampaio University Center, Juazeiro do Norte, CEP 63040-405, BrazilDoctor Leão Sampaio University Center, Juazeiro do Norte, CEP 63040-405, BrazilDoctor Leão Sampaio University Center, Juazeiro do Norte, CEP 63040-405, BrazilDoctor Leão Sampaio University Center, Juazeiro do Norte, CEP 63040-405, BrazilDoctor Leão Sampaio University Center, Juazeiro do Norte, CEP 63040-405, BrazilDoctor Leão Sampaio University Center, Juazeiro do Norte, CEP 63040-405, BrazilFederal University of Ceara, CEP 60020-181, Fortaleza, BrazilFederal University of Cariri, Crato, CEP 63130-025, BrazilDoctor Leão Sampaio University Center, Juazeiro do Norte, CEP 63040-405, BrazilBackground and objectives: Given the difficulty in establishing an ideal method that can successfully extract bacterial deoxyribonucleic acid (DNA) in Gram-negative bacteria, the objective of this work was to compare protocols for the extraction of bacterial DNA and to report the more practical, faster, and less costly ones. Methods: Bacterial species used were Escherichia coli, provided by Dr. Leão Sampaio University Center. The methods used to extract bacterial DNA were: sodium dodecyl sulfate (SDS), salting-out, cetyltrimethylammonium bromide (CTAB)/Phenol-Chloroform, and commercial kit for DNA isolation from Promega. The extracted DNA by all the methods was detected and assessed by the agarose gel electrophoresis. Results: The best result was that obtained by the SDS method, it showed the greatest advantage for bringing speed, low cost, and good concentration of the extracted material. The other methods showed low-quality DNA, probably due to the presence of large amounts of proteins in the cell wall of E. coli interfering with the quality of the samples. Conclusions: It was concluded that the SDS method can provide better DNA in terms of quality, which is preferred for the amplification by the polymerase (PCR) chain reaction. This will be helpful for diagnostic microbiologists because of the low cost and good quality of the extracted DNA.https://bjbabs.org/index.php/bjbabs/article/view/123dna extractionelectrophoresisescherichia colisds |
| spellingShingle | Oslaene Alves de Brito Francisca Alves dos Santos Tassia Thais Al Yafawi Cícero Roberto Nascimento Saraiva Rakel Olinda Macedo da Silva Lívia Maria Garcia Leandro Pedro Everson Alexandre de Aquino Dárcio Luiz de Sousa Júnior Maria Karollyna do Nascimento Silva Leandro Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis Baghdad Journal of Biochemistry and Applied Biological Sciences dna extraction electrophoresis escherichia coli sds |
| title | Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis |
| title_full | Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis |
| title_fullStr | Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis |
| title_full_unstemmed | Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis |
| title_short | Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis |
| title_sort | comparing protocols of dna extraction from escherichia coli analysis of purity and concentration by gel electrophoresis |
| topic | dna extraction electrophoresis escherichia coli sds |
| url | https://bjbabs.org/index.php/bjbabs/article/view/123 |
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