Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA.
The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/uL (7 days in transport buffer) to 702.00 ng/uL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/uL (30 days in silica gel) to 2,850.00 ng/uL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies.
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Format: | Artigo de periódico biblioteca |
Language: | English eng |
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2020-01-21
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Subjects: | Leaflet, Hojas, Extracción, Maceración, Embrapa Acre, Rio Branco, Acre, Amazônia Ocidental, Western Amazon, Amazonia Occidental., Açaí, Extração, DNA, Campo Experimental., Maceração, Folha, Extraction, Leaves, Euterpe precatoria, Maceration., |
Online Access: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1119144 |
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dig-alice-doc-11191442020-01-22T00:38:02Z Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA. AZÊVEDO, H. S. F. da S. RUFINO, P. B. AZEVEDO, J. M. A. de SILVA, L. M. da WADT, L. H. de O. CAMPOS, T. de Hellen Sandra Freires da Silva Azêvedo, Fundação Oswaldo Cruz (FIOCRUZ), Rede Bionorte; Polinar Bandeira Rufino, Faculdade Meta (FAMETA); José Marlo Araújo de Azevedo, Instituto Federal do Acre (IFAC); LUCIELIO MANOEL DA SILVA, CPAF-AC; LUCIA HELENA DE OLIVEIRA WADT, CPAF-RO; TATIANA DE CAMPOS, CPAF-AC. Leaflet Hojas Extracción Maceración Embrapa Acre Rio Branco Acre Amazônia Ocidental Western Amazon Amazonia Occidental. Açaí Extração DNA Campo Experimental. Maceração Folha Extraction Leaves Euterpe precatoria Maceration. The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/uL (7 days in transport buffer) to 702.00 ng/uL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/uL (30 days in silica gel) to 2,850.00 ng/uL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies. 2020-01-22T00:37:56Z 2020-01-22T00:37:56Z 2020-01-21 2019 2020-01-27T11:11:11Z Artigo de periódico Bioscience Journal, v. 35, n. 4, p. 1188-1197, jul./ago. 2019. 1981-3163 http://www.alice.cnptia.embrapa.br/alice/handle/doc/1119144 10.14393/BJ-v35n4a2019-42190 en eng openAccess |
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Leaflet Hojas Extracción Maceración Embrapa Acre Rio Branco Acre Amazônia Ocidental Western Amazon Amazonia Occidental. Açaí Extração DNA Campo Experimental. Maceração Folha Extraction Leaves Euterpe precatoria Maceration. Leaflet Hojas Extracción Maceración Embrapa Acre Rio Branco Acre Amazônia Ocidental Western Amazon Amazonia Occidental. Açaí Extração DNA Campo Experimental. Maceração Folha Extraction Leaves Euterpe precatoria Maceration. |
spellingShingle |
Leaflet Hojas Extracción Maceración Embrapa Acre Rio Branco Acre Amazônia Ocidental Western Amazon Amazonia Occidental. Açaí Extração DNA Campo Experimental. Maceração Folha Extraction Leaves Euterpe precatoria Maceration. Leaflet Hojas Extracción Maceración Embrapa Acre Rio Branco Acre Amazônia Ocidental Western Amazon Amazonia Occidental. Açaí Extração DNA Campo Experimental. Maceração Folha Extraction Leaves Euterpe precatoria Maceration. AZÊVEDO, H. S. F. da S. RUFINO, P. B. AZEVEDO, J. M. A. de SILVA, L. M. da WADT, L. H. de O. CAMPOS, T. de Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA. |
description |
The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/uL (7 days in transport buffer) to 702.00 ng/uL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/uL (30 days in silica gel) to 2,850.00 ng/uL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies. |
author2 |
Hellen Sandra Freires da Silva Azêvedo, Fundação Oswaldo Cruz (FIOCRUZ), Rede Bionorte; Polinar Bandeira Rufino, Faculdade Meta (FAMETA); José Marlo Araújo de Azevedo, Instituto Federal do Acre (IFAC); LUCIELIO MANOEL DA SILVA, CPAF-AC; LUCIA HELENA DE OLIVEIRA WADT, CPAF-RO; TATIANA DE CAMPOS, CPAF-AC. |
author_facet |
Hellen Sandra Freires da Silva Azêvedo, Fundação Oswaldo Cruz (FIOCRUZ), Rede Bionorte; Polinar Bandeira Rufino, Faculdade Meta (FAMETA); José Marlo Araújo de Azevedo, Instituto Federal do Acre (IFAC); LUCIELIO MANOEL DA SILVA, CPAF-AC; LUCIA HELENA DE OLIVEIRA WADT, CPAF-RO; TATIANA DE CAMPOS, CPAF-AC. AZÊVEDO, H. S. F. da S. RUFINO, P. B. AZEVEDO, J. M. A. de SILVA, L. M. da WADT, L. H. de O. CAMPOS, T. de |
format |
Artigo de periódico |
topic_facet |
Leaflet Hojas Extracción Maceración Embrapa Acre Rio Branco Acre Amazônia Ocidental Western Amazon Amazonia Occidental. Açaí Extração DNA Campo Experimental. Maceração Folha Extraction Leaves Euterpe precatoria Maceration. |
author |
AZÊVEDO, H. S. F. da S. RUFINO, P. B. AZEVEDO, J. M. A. de SILVA, L. M. da WADT, L. H. de O. CAMPOS, T. de |
author_sort |
AZÊVEDO, H. S. F. da S. |
title |
Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA. |
title_short |
Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA. |
title_full |
Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA. |
title_fullStr |
Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA. |
title_full_unstemmed |
Preservation and maceration of Amazon açai leaflet tissue to obtain genomic DNA. |
title_sort |
preservation and maceration of amazon açai leaflet tissue to obtain genomic dna. |
publishDate |
2020-01-21 |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1119144 |
work_keys_str_mv |
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_version_ |
1756026756598333440 |