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...
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| Format: | Article |
| Language: | English |
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Universidade Federal de Uberlândia
2019-06-01
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| Series: | Bioscience Journal |
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| Online Access: | https://seer-dev.ufu.br/index.php/biosciencejournal/article/view/42190 |
| _version_ | 1818157021652844544 |
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| author | Hellen Sandra Freires da Silva Azêvedo Polinar Bandeira Rufino José Marlo Araújo de Azevedo Luciélio Manoel da Silva Lúcia Helena de Oliveira Wadt Tatiana de Campos |
| author_facet | Hellen Sandra Freires da Silva Azêvedo Polinar Bandeira Rufino José Marlo Araújo de Azevedo Luciélio Manoel da Silva Lúcia Helena de Oliveira Wadt Tatiana de Campos |
| author_sort | Hellen Sandra Freires da Silva Azêvedo |
| collection | DOAJ |
| 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/µL (7 days in transport buffer) to 702.00 ng/µL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µL (30 days in silica gel) to 2,850.00 ng/µL (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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| first_indexed | 2024-12-11T15:07:34Z |
| format | Article |
| id | doaj.art-0501af48d43a4724bd26d6c6902d7741 |
| institution | Directory Open Access Journal |
| issn | 1981-3163 |
| language | English |
| last_indexed | 2024-12-11T15:07:34Z |
| publishDate | 2019-06-01 |
| publisher | Universidade Federal de Uberlândia |
| record_format | Article |
| series | Bioscience Journal |
| spelling | doaj.art-0501af48d43a4724bd26d6c6902d77412022-12-22T01:00:50ZengUniversidade Federal de UberlândiaBioscience Journal1981-31632019-06-01354Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNAHellen Sandra Freires da Silva Azêvedo0Polinar Bandeira Rufino1José Marlo Araújo de Azevedo2Luciélio Manoel da Silva3Lúcia Helena de Oliveira Wadt4Tatiana de Campos5Fundação Oswaldo Cruz (FIOCRUZ).Faculdade Meta (FAMETA)Instituto Federal do Acre (IFAC)Empresa Brasileira de Pesquisa Agropecuária, Embrapa AcreEmpresa Brasileira de Pesquisa Agropecuária, Embrapa RondôniaEmpresa Brasileira de Pesquisa Agropecuária, Embrapa Acre 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/µL (7 days in transport buffer) to 702.00 ng/µL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µL (30 days in silica gel) to 2,850.00 ng/µL (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. https://seer-dev.ufu.br/index.php/biosciencejournal/article/view/42190Açaí-solteiroCTAB methodDNA isolationEuterpe precatoria |
| spellingShingle | Hellen Sandra Freires da Silva Azêvedo Polinar Bandeira Rufino José Marlo Araújo de Azevedo Luciélio Manoel da Silva Lúcia Helena de Oliveira Wadt Tatiana de Campos Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA Bioscience Journal Açaí-solteiro CTAB method DNA isolation Euterpe precatoria |
| title | 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_short | Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA |
| title_sort | preservation and maceration of amazon acai leaflet tissue to obtain genomic dna |
| topic | Açaí-solteiro CTAB method DNA isolation Euterpe precatoria |
| url | https://seer-dev.ufu.br/index.php/biosciencejournal/article/view/42190 |
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