Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2022 |
| Outros Autores: | , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Acta Scientiarum Biological Sciences |
| Texto Completo: | http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/59272 |
Resumo: | DNA extraction is usually the first step to perform molecular studies. This process can be nonviable due to genomic DNA (gDNA) extraction commercial kits prices. Furthermore, available DNA extraction protocols generally have high specificity, limiting their use to specific sources of biological material. In order to reduce costs, optimize time and laboratory logistics, besides to demonstrate a versatile protocol, the present study worked on an efficient DNA extraction protocol from somatic and non-somatic cells, using biological material from sheep as a model. For that, gDNA was extracted from whole blood, spermatozoa, and hair bulb cells, collected from three adult sheep, transported at 5ºC and stored at -20ºC until lab procedures. After extraction, gDNA concentration and purity were evaluated in a nano spectrophotometer. gDNA concentration from whole blood was greater (p < 0.05) than extracted from hair bulb cells, which in turn was superior (p < 0.05) than in spermatozoa. Also, gDNA from whole blood and, followed by, sperm showed greater (p < 0.05) purity when compared to gDNA of hair bulb cells. Adapting a gDNA extraction protocol, originally developed for bovine whole blood, enabled to obtain and isolate gDNA in different nucleated sheep cells |
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Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement.Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement.DNA extraction is usually the first step to perform molecular studies. This process can be nonviable due to genomic DNA (gDNA) extraction commercial kits prices. Furthermore, available DNA extraction protocols generally have high specificity, limiting their use to specific sources of biological material. In order to reduce costs, optimize time and laboratory logistics, besides to demonstrate a versatile protocol, the present study worked on an efficient DNA extraction protocol from somatic and non-somatic cells, using biological material from sheep as a model. For that, gDNA was extracted from whole blood, spermatozoa, and hair bulb cells, collected from three adult sheep, transported at 5ºC and stored at -20ºC until lab procedures. After extraction, gDNA concentration and purity were evaluated in a nano spectrophotometer. gDNA concentration from whole blood was greater (p < 0.05) than extracted from hair bulb cells, which in turn was superior (p < 0.05) than in spermatozoa. Also, gDNA from whole blood and, followed by, sperm showed greater (p < 0.05) purity when compared to gDNA of hair bulb cells. Adapting a gDNA extraction protocol, originally developed for bovine whole blood, enabled to obtain and isolate gDNA in different nucleated sheep cellsDNA extraction is usually the first step to perform molecular studies. This process can be nonviable due to genomic DNA (gDNA) extraction commercial kits prices. Furthermore, available DNA extraction protocols generally have high specificity, limiting their use to specific sources of biological material. In order to reduce costs, optimize time and laboratory logistics, besides to demonstrate a versatile protocol, the present study worked on an efficient DNA extraction protocol from somatic and non-somatic cells, using biological material from sheep as a model. For that, gDNA was extracted from whole blood, spermatozoa, and hair bulb cells, collected from three adult sheep, transported at 5ºC and stored at -20ºC until lab procedures. After extraction, gDNA concentration and purity were evaluated in a nano spectrophotometer. gDNA concentration from whole blood was greater (p < 0.05) than extracted from hair bulb cells, which in turn was superior (p < 0.05) than in spermatozoa. Also, gDNA from whole blood and, followed by, sperm showed greater (p < 0.05) purity when compared to gDNA of hair bulb cells. Adapting a gDNA extraction protocol, originally developed for bovine whole blood, enabled to obtain and isolate gDNA in different nucleated sheep cellsUniversidade Estadual De Maringá2022-07-27info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/5927210.4025/actascibiolsci.v44i1.59272Acta Scientiarum. Biological Sciences; Vol 44 (2022): Publicação contínua; e59272Acta Scientiarum. Biological Sciences; v. 44 (2022): Publicação contínua; e592721807-863X1679-9283reponame:Acta Scientiarum Biological Sciencesinstname:Universidade Estadual de Maringá (UEM)instacron:UEMenghttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/59272/751375154601Copyright (c) 2022 Acta Scientiarum. Biological Scienceshttp://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccess Santos, André Luiz da ConceiçãoSilva, Wilson Romão Toledo da Ferreira, Heder Nunes Fraga Junior, Antonio Matos Batista, Marcus Vinicius de Aragão Velarde, Jonatan Mikhail Del Solar 2022-08-22T16:54:30Zoai:periodicos.uem.br/ojs:article/59272Revistahttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSciPUBhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/oai||actabiol@uem.br1807-863X1679-9283opendoar:2022-08-22T16:54:30Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM)false |
| dc.title.none.fl_str_mv |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells |
| title |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells |
| spellingShingle |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells Santos, André Luiz da Conceição Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement. Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement. |
| title_short |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells |
| title_full |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells |
| title_fullStr |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells |
| title_full_unstemmed |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells |
| title_sort |
Efficient, fast and low-cost strategies for DNA extraction from different nucleated sheep cells |
| author |
Santos, André Luiz da Conceição |
| author_facet |
Santos, André Luiz da Conceição Silva, Wilson Romão Toledo da Ferreira, Heder Nunes Fraga Junior, Antonio Matos Batista, Marcus Vinicius de Aragão Velarde, Jonatan Mikhail Del Solar |
| author_role |
author |
| author2 |
Silva, Wilson Romão Toledo da Ferreira, Heder Nunes Fraga Junior, Antonio Matos Batista, Marcus Vinicius de Aragão Velarde, Jonatan Mikhail Del Solar |
| author2_role |
author author author author author |
| dc.contributor.author.fl_str_mv |
Santos, André Luiz da Conceição Silva, Wilson Romão Toledo da Ferreira, Heder Nunes Fraga Junior, Antonio Matos Batista, Marcus Vinicius de Aragão Velarde, Jonatan Mikhail Del Solar |
| dc.subject.por.fl_str_mv |
Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement. Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement. |
| topic |
Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement. Biotechnology; DNA extraction; hair bulb; spermatozoa; blood; genetic improvement. |
| description |
DNA extraction is usually the first step to perform molecular studies. This process can be nonviable due to genomic DNA (gDNA) extraction commercial kits prices. Furthermore, available DNA extraction protocols generally have high specificity, limiting their use to specific sources of biological material. In order to reduce costs, optimize time and laboratory logistics, besides to demonstrate a versatile protocol, the present study worked on an efficient DNA extraction protocol from somatic and non-somatic cells, using biological material from sheep as a model. For that, gDNA was extracted from whole blood, spermatozoa, and hair bulb cells, collected from three adult sheep, transported at 5ºC and stored at -20ºC until lab procedures. After extraction, gDNA concentration and purity were evaluated in a nano spectrophotometer. gDNA concentration from whole blood was greater (p < 0.05) than extracted from hair bulb cells, which in turn was superior (p < 0.05) than in spermatozoa. Also, gDNA from whole blood and, followed by, sperm showed greater (p < 0.05) purity when compared to gDNA of hair bulb cells. Adapting a gDNA extraction protocol, originally developed for bovine whole blood, enabled to obtain and isolate gDNA in different nucleated sheep cells |
| publishDate |
2022 |
| dc.date.none.fl_str_mv |
2022-07-27 |
| dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/59272 10.4025/actascibiolsci.v44i1.59272 |
| url |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/59272 |
| identifier_str_mv |
10.4025/actascibiolsci.v44i1.59272 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/59272/751375154601 |
| dc.rights.driver.fl_str_mv |
Copyright (c) 2022 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
| rights_invalid_str_mv |
Copyright (c) 2022 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
| publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
| dc.source.none.fl_str_mv |
Acta Scientiarum. Biological Sciences; Vol 44 (2022): Publicação contínua; e59272 Acta Scientiarum. Biological Sciences; v. 44 (2022): Publicação contínua; e59272 1807-863X 1679-9283 reponame:Acta Scientiarum Biological Sciences instname:Universidade Estadual de Maringá (UEM) instacron:UEM |
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Universidade Estadual de Maringá (UEM) |
| instacron_str |
UEM |
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UEM |
| reponame_str |
Acta Scientiarum Biological Sciences |
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Acta Scientiarum Biological Sciences |
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Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM) |
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||actabiol@uem.br |
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1799317397985820672 |