Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae)
Autor(a) principal: | |
---|---|
Data de Publicação: | 2014 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/16693 |
Resumo: | Nectandra megapotamica (Spreng.) Mez is a native tree species of ecological and pharmacological importance, due to its great potential for the production of drugs. The selection of genetically superior individuals is a feasible alternative to leverage the studies regarding this potential. However, because it is a species that produces large amounts of secondary metabolites, the isolation of genomic DNA in sufficient quantities and of good quality, is crucial step for the development of any technique for direct analysis of deoxyribonucleic acid. Considering the above, the overall objective of this study was to define a protocol for DNA extraction, specific to Nectandra megapotamica (Spreng.) Mez. Able to obtain samples of quality and quantity for further applications. The foliar samples dried at room temperature, dried in oven at 40 °C and dried in a microwave oven, plus four DNA extraction protocols were tested (Dellaporta, Ferreira and Grattapaglia, Khanuja e Mazza and Bittencourt). The ratios A260/A230, A260/A280, the concentration of DNA obtained, DNA integrity, functionality DNA via digestion with Hind III enzyme and the correlation between the expected absorbance curve for a DNA sample and the observed curve were evaluated. For the types of foliar samples was found that it is possible to extract DNA from both. However, only dried at room temperature samples showed results for quality and quantity as expected. In the analysis of different extraction protocols, Mazza and Bittencourt and Ferreira and Grattapaglia showed the best results for DNA concentration. However, these exhibited high levels of contamination, verified by the gelatinous aspect and brown coloration the end of the procedure, confirming the results indicated by ratios. Correlation analysis of the absorbance curve for these two protocols showed that the absorbance peaks were 220 nm and 245 nm, respectively, indicating high levels of phenolic compounds or polysaccharides in the samples. The Dellaporta protocol, showed the best results for two reasons, one concentration of intermediate DNA, the highest correlation between expected and observed DNA curve. In addition the extracted DNA was digested by the enzyme Hind III, which was not observed for other protocols. With the intention to optimize the Dellaporta protocol, tests were performed with different concentrations of SDS (10, 15, 20 and 25%), PVP (0, 1, 2 and 3%), potassium acetate (2,5; 5; 7,5 and 10 M) and NaCl in the elution buffer (0, 0,5; 1 and 1,5 M), wherein the DNA concentration, the A260/A280 and A230/A230 ratio and the correlation between the absorbance curves were evaluated. Tests with NaCl in the elution buffer and Potassium Acetate have not resulted significant gains, indicating their respective use according the original protocol. For SDS, concentrations of 15 and 25% the greatest amount of extracted DNA. However, 15% presented a ratio A260/A230 lower than expected. Regarding PVP, in absence and presence of 1% concentration presented very close results, however, the latter showed greater correlation between the absorbance curves. The combined analysis of all variables, indicated that the use of 25% SDS and 1% PVP in Dellaporta protocol, contributes to improve concentration and quality of isolated DNA. |
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2019-05-29T14:26:37Z2019-05-29T14:26:37Z2014-02-28http://repositorio.ufsm.br/handle/1/16693Nectandra megapotamica (Spreng.) Mez is a native tree species of ecological and pharmacological importance, due to its great potential for the production of drugs. The selection of genetically superior individuals is a feasible alternative to leverage the studies regarding this potential. However, because it is a species that produces large amounts of secondary metabolites, the isolation of genomic DNA in sufficient quantities and of good quality, is crucial step for the development of any technique for direct analysis of deoxyribonucleic acid. Considering the above, the overall objective of this study was to define a protocol for DNA extraction, specific to Nectandra megapotamica (Spreng.) Mez. Able to obtain samples of quality and quantity for further applications. The foliar samples dried at room temperature, dried in oven at 40 °C and dried in a microwave oven, plus four DNA extraction protocols were tested (Dellaporta, Ferreira and Grattapaglia, Khanuja e Mazza and Bittencourt). The ratios A260/A230, A260/A280, the concentration of DNA obtained, DNA integrity, functionality DNA via digestion with Hind III enzyme and the correlation between the expected absorbance curve for a DNA sample and the observed curve were evaluated. For the types of foliar samples was found that it is possible to extract DNA from both. However, only dried at room temperature samples showed results for quality and quantity as expected. In the analysis of different extraction protocols, Mazza and Bittencourt and Ferreira and Grattapaglia showed the best results for DNA concentration. However, these exhibited high levels of contamination, verified by the gelatinous aspect and brown coloration the end of the procedure, confirming the results indicated by ratios. Correlation analysis of the absorbance curve for these two protocols showed that the absorbance peaks were 220 nm and 245 nm, respectively, indicating high levels of phenolic compounds or polysaccharides in the samples. The Dellaporta protocol, showed the best results for two reasons, one concentration of intermediate DNA, the highest correlation between expected and observed DNA curve. In addition the extracted DNA was digested by the enzyme Hind III, which was not observed for other protocols. With the intention to optimize the Dellaporta protocol, tests were performed with different concentrations of SDS (10, 15, 20 and 25%), PVP (0, 1, 2 and 3%), potassium acetate (2,5; 5; 7,5 and 10 M) and NaCl in the elution buffer (0, 0,5; 1 and 1,5 M), wherein the DNA concentration, the A260/A280 and A230/A230 ratio and the correlation between the absorbance curves were evaluated. Tests with NaCl in the elution buffer and Potassium Acetate have not resulted significant gains, indicating their respective use according the original protocol. For SDS, concentrations of 15 and 25% the greatest amount of extracted DNA. However, 15% presented a ratio A260/A230 lower than expected. Regarding PVP, in absence and presence of 1% concentration presented very close results, however, the latter showed greater correlation between the absorbance curves. The combined analysis of all variables, indicated that the use of 25% SDS and 1% PVP in Dellaporta protocol, contributes to improve concentration and quality of isolated DNA.Nectandra megapotamica (Spreng.) Mez é uma espécie florestal nativa de importância ecológica e farmacológica, devido ao seu grande potencial de produção de fármacos. A seleção de indivíduos geneticamente superiores é uma alternativa viável para alavancar os estudos referentes a esse potencial. No entanto, por se tratar de uma espécie que produz grandes quantidades de metabólitos secundários, o isolamento de DNA genômico em quantidades suficientes e de boa qualidade, é um passo crucial para o desenvolvimento de qualquer técnica de análise direta do ácido desoxirribonucleico. Considerando o exposto, o objetivo geral do presente estudo foi definir um protocolo de extração de DNA, específico para Nectandra megapotamica (Spreng.) Mez., capaz de obter amostras de qualidade e quantidade para aplicações posteriores. Foram testadas amostras foliares secas à temperatura ambiente, secas em estufa à 40 °C e secas em forno micro-ondas, além de quatro protocolos de extração de DNA (Dellaporta, Ferreira e Grattapaglia, Khanuja e Mazza e Bittencourt). Foram avaliadas as razões A260/A230, A260/A280, a concentração de DNA obtida, a integridade do DNA, a funcionalidade do DNA via digestão pela enzima Hind III e a correlação entre a curva de absorvância esperada para uma amostra de DNA e a curva observada. Para os tipos de amostras foliares, foi verificado que é possível extrair DNA dos três tipos de amostras foliares. Porém, apenas as amostras secas em temperatura ambiente apresentaram resultados para qualidade e quantidade dentro do esperado. Na análise dos diferentes protocolos de extração, Mazza e Bittencourt e Ferreira e Grattapaglia apresentaram os melhores resultados para concentração de DNA. No entanto, esses exibiram elevados teores de contaminação, verificado pelo aspecto gelatinoso e coloração castanha ao final do procedimento, comprovando o indicado pelos resultados das razões. A análise da correlação da curva de absorvância para esses dois protocolos, revelou que os picos de absorvância foram em 220 nm e 245 nm, respectivamente, indicando altos teores compostos fenólicos ou polissacarídeos nas amostras. O protocolo Dellaporta apresentou os melhores resultados para as duas razões, uma concentração de DNA intermediária, a maior correlação entre a curva de DNA esperada e a observada, além do DNA extraído ter sido digerido pela enzima Hind III, o que não foi verificado para os demais protocolos. Com o intuito de otimizar o protocolo Dellaporta, foram realizados testes com diferentes concentrações de SDS (10, 15, 20 e 25%), PVP (0, 1, 2 e 3%), Acetato de Potássio (2,5; 5; 7,5 e 10 M) e NaCl no tampão de eluição (0; 0,5; 1 e 1,5 M), em que foram avaliadas a concentração de DNA, a razão A260/A280 e A230/A230 e correlação entre as curvas de absorvância. Os testes com NaCl no tampão de eluição e Acetato de Potássio, não resultaram ganhos significativos, indicando os seus respectivos usos conforme o protocolo original. Para SDS, as concentrações de 15 e 25% extraíram a maior quantidade de DNA. Porém, 15% apresentou uma razão A260/A230 abaixo do esperado. Com relação ao PVP, a ausência e a presença na concentração de 1% apresentaram resultados muito próximos, porém, o último apresentou uma maior correlação entre as curvas de absorvância. A análise conjunta de todas as variáveis, indica que o uso de SDS 25% e PVP 1% no protocolo Dellaporta, contribuiram para melhorar a concentração e a qualidade do DNA isolado.porUniversidade Federal de Santa MariaCentro de Ciências RuraisPrograma de Pós-Graduação em Engenharia FlorestalUFSMBrasilRecursos Florestais e Engenharia FlorestalAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessCanela-pretaCTABSDSMetabólitos secundáriosPolissacarídeosEnzimas de restriçãoSecondary metabolitesPolysaccharidesRestriction enzymesCNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTALIsolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae)Genomic DNA isolation from leaves samples of Nectandra megapotamica (Spreng.) Mez. (Lauraceae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisReiniger, Lia Rejane Silveirahttp://lattes.cnpq.br/5739294882585391Stefenon, Valdir Marcoshttp://lattes.cnpq.br/6868213051236665Stefenon, Valdir Marcoshttp://lattes.cnpq.br/6868213051236665Bevilacqua, Caroline Borgeshttp://lattes.cnpq.br/3635265675594303http://lattes.cnpq.br/2680625582907390Costa, Leonardo Severo da500200000003600500809860fd-9f1f-4d0e-81fb-36d7e9781d1c9583d8c5-8c6a-4d1d-864b-8bfd70c695190285882e-f5b8-432f-ad65-54769c51a1551c88dca6-51c2-497a-95a8-36df433f22c7120fe11e-ba09-4c03-ba31-100bdf7e46aareponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALDIS_PPGEF_2014_COSTA_LEONARDO.pdfDIS_PPGEF_2014_COSTA_LEONARDO.pdfDissertação de Mestradoapplication/pdf3889138http://repositorio.ufsm.br/bitstream/1/16693/1/DIS_PPGEF_2014_COSTA_LEONARDO.pdf91c8bee86fb2a7504de83042b1eed754MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805http://repositorio.ufsm.br/bitstream/1/16693/2/license_rdf4460e5956bc1d1639be9ae6146a50347MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81956http://repositorio.ufsm.br/bitstream/1/16693/3/license.txt2f0571ecee68693bd5cd3f17c1e075dfMD53TEXTDIS_PPGEF_2014_COSTA_LEONARDO.pdf.txtDIS_PPGEF_2014_COSTA_LEONARDO.pdf.txtExtracted texttext/plain162860http://repositorio.ufsm.br/bitstream/1/16693/4/DIS_PPGEF_2014_COSTA_LEONARDO.pdf.txtd1ac724d510709606995a0cbe2fa1571MD54THUMBNAILDIS_PPGEF_2014_COSTA_LEONARDO.pdf.jpgDIS_PPGEF_2014_COSTA_LEONARDO.pdf.jpgIM Thumbnailimage/jpeg5022http://repositorio.ufsm.br/bitstream/1/16693/5/DIS_PPGEF_2014_COSTA_LEONARDO.pdf.jpg85617f079a4d0f8784391505df681cdbMD551/166932022-05-18 11:28:05.739oai:repositorio.ufsm.br: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ório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132022-05-18T14:28:05Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
dc.title.alternative.eng.fl_str_mv |
Genomic DNA isolation from leaves samples of Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
title |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
spellingShingle |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) Costa, Leonardo Severo da Canela-preta CTAB SDS Metabólitos secundários Polissacarídeos Enzimas de restrição Secondary metabolites Polysaccharides Restriction enzymes CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL |
title_short |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
title_full |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
title_fullStr |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
title_full_unstemmed |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
title_sort |
Isolamento de DNA genômico em amostras foliares de Nectandra megapotamica (Spreng.) Mez. (Lauraceae) |
author |
Costa, Leonardo Severo da |
author_facet |
Costa, Leonardo Severo da |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Reiniger, Lia Rejane Silveira |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5739294882585391 |
dc.contributor.advisor-co1.fl_str_mv |
Stefenon, Valdir Marcos |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/6868213051236665 |
dc.contributor.referee1.fl_str_mv |
Stefenon, Valdir Marcos |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/6868213051236665 |
dc.contributor.referee2.fl_str_mv |
Bevilacqua, Caroline Borges |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/3635265675594303 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2680625582907390 |
dc.contributor.author.fl_str_mv |
Costa, Leonardo Severo da |
contributor_str_mv |
Reiniger, Lia Rejane Silveira Stefenon, Valdir Marcos Stefenon, Valdir Marcos Bevilacqua, Caroline Borges |
dc.subject.por.fl_str_mv |
Canela-preta CTAB SDS Metabólitos secundários Polissacarídeos Enzimas de restrição |
topic |
Canela-preta CTAB SDS Metabólitos secundários Polissacarídeos Enzimas de restrição Secondary metabolites Polysaccharides Restriction enzymes CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL |
dc.subject.eng.fl_str_mv |
Secondary metabolites Polysaccharides Restriction enzymes |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::RECURSOS FLORESTAIS E ENGENHARIA FLORESTAL |
description |
Nectandra megapotamica (Spreng.) Mez is a native tree species of ecological and pharmacological importance, due to its great potential for the production of drugs. The selection of genetically superior individuals is a feasible alternative to leverage the studies regarding this potential. However, because it is a species that produces large amounts of secondary metabolites, the isolation of genomic DNA in sufficient quantities and of good quality, is crucial step for the development of any technique for direct analysis of deoxyribonucleic acid. Considering the above, the overall objective of this study was to define a protocol for DNA extraction, specific to Nectandra megapotamica (Spreng.) Mez. Able to obtain samples of quality and quantity for further applications. The foliar samples dried at room temperature, dried in oven at 40 °C and dried in a microwave oven, plus four DNA extraction protocols were tested (Dellaporta, Ferreira and Grattapaglia, Khanuja e Mazza and Bittencourt). The ratios A260/A230, A260/A280, the concentration of DNA obtained, DNA integrity, functionality DNA via digestion with Hind III enzyme and the correlation between the expected absorbance curve for a DNA sample and the observed curve were evaluated. For the types of foliar samples was found that it is possible to extract DNA from both. However, only dried at room temperature samples showed results for quality and quantity as expected. In the analysis of different extraction protocols, Mazza and Bittencourt and Ferreira and Grattapaglia showed the best results for DNA concentration. However, these exhibited high levels of contamination, verified by the gelatinous aspect and brown coloration the end of the procedure, confirming the results indicated by ratios. Correlation analysis of the absorbance curve for these two protocols showed that the absorbance peaks were 220 nm and 245 nm, respectively, indicating high levels of phenolic compounds or polysaccharides in the samples. The Dellaporta protocol, showed the best results for two reasons, one concentration of intermediate DNA, the highest correlation between expected and observed DNA curve. In addition the extracted DNA was digested by the enzyme Hind III, which was not observed for other protocols. With the intention to optimize the Dellaporta protocol, tests were performed with different concentrations of SDS (10, 15, 20 and 25%), PVP (0, 1, 2 and 3%), potassium acetate (2,5; 5; 7,5 and 10 M) and NaCl in the elution buffer (0, 0,5; 1 and 1,5 M), wherein the DNA concentration, the A260/A280 and A230/A230 ratio and the correlation between the absorbance curves were evaluated. Tests with NaCl in the elution buffer and Potassium Acetate have not resulted significant gains, indicating their respective use according the original protocol. For SDS, concentrations of 15 and 25% the greatest amount of extracted DNA. However, 15% presented a ratio A260/A230 lower than expected. Regarding PVP, in absence and presence of 1% concentration presented very close results, however, the latter showed greater correlation between the absorbance curves. The combined analysis of all variables, indicated that the use of 25% SDS and 1% PVP in Dellaporta protocol, contributes to improve concentration and quality of isolated DNA. |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-02-28 |
dc.date.accessioned.fl_str_mv |
2019-05-29T14:26:37Z |
dc.date.available.fl_str_mv |
2019-05-29T14:26:37Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/16693 |
url |
http://repositorio.ufsm.br/handle/1/16693 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
500200000003 |
dc.relation.confidence.fl_str_mv |
600 500 |
dc.relation.authority.fl_str_mv |
809860fd-9f1f-4d0e-81fb-36d7e9781d1c 9583d8c5-8c6a-4d1d-864b-8bfd70c69519 0285882e-f5b8-432f-ad65-54769c51a155 1c88dca6-51c2-497a-95a8-36df433f22c7 120fe11e-ba09-4c03-ba31-100bdf7e46aa |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Engenharia Florestal |
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UFSM |
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Brasil |
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Recursos Florestais e Engenharia Florestal |
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Universidade Federal de Santa Maria Centro de Ciências Rurais |
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Universidade Federal de Santa Maria (UFSM) |
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UFSM |
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UFSM |
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