A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment
ABSTRACT Many tropical trees have high canopies and their leaves are not accessible. Thus, the use of tissue from a more accessible organ (cambium) for DNA extraction may be an alternative for molecular studies. We adapted a feasible methodology for extracting genomic DNA from cambium tissue harvested in the field for the assessment with PCR. We tested three storage conditions (two buffers and a silica gel) and four periods of time after harvest. We used previously described protocols and tested them on three species that occur in Amazonian forests and other biomes: Anadenanthera peregrina var. peregrina, Cedrela fissilis, and Ceiba speciosa. Our protocol obtained suitable PCR-grade genomic DNA for DNA sequencing and microsatellite genotyping. We recommend the use of silica for long-term storage and the buffer with ascorbic acid for short-term storage.
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Instituto Nacional de Pesquisas da Amazônia
2020
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oai:scielo:S0044-596720200004003352020-12-04A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessmentMANGARAVITE,ÉricaTERRA,VanessaHATTORI,Eric Koiti OkiyamaDAL’SASSO,Thaís Carolina da SilvaBHERING,Leonardo LopesOLIVEIRA,Luiz Orlando de ascorbic acid dithiothreitol DNA isolation ABSTRACT Many tropical trees have high canopies and their leaves are not accessible. Thus, the use of tissue from a more accessible organ (cambium) for DNA extraction may be an alternative for molecular studies. We adapted a feasible methodology for extracting genomic DNA from cambium tissue harvested in the field for the assessment with PCR. We tested three storage conditions (two buffers and a silica gel) and four periods of time after harvest. We used previously described protocols and tested them on three species that occur in Amazonian forests and other biomes: Anadenanthera peregrina var. peregrina, Cedrela fissilis, and Ceiba speciosa. Our protocol obtained suitable PCR-grade genomic DNA for DNA sequencing and microsatellite genotyping. We recommend the use of silica for long-term storage and the buffer with ascorbic acid for short-term storage.info:eu-repo/semantics/openAccessInstituto Nacional de Pesquisas da AmazôniaActa Amazonica v.50 n.4 20202020-12-01info:eu-repo/semantics/reporttext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0044-59672020000400335en10.1590/1809-4392202001571 |
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MANGARAVITE,Érica TERRA,Vanessa HATTORI,Eric Koiti Okiyama DAL’SASSO,Thaís Carolina da Silva BHERING,Leonardo Lopes OLIVEIRA,Luiz Orlando de |
| spellingShingle |
MANGARAVITE,Érica TERRA,Vanessa HATTORI,Eric Koiti Okiyama DAL’SASSO,Thaís Carolina da Silva BHERING,Leonardo Lopes OLIVEIRA,Luiz Orlando de A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment |
| author_facet |
MANGARAVITE,Érica TERRA,Vanessa HATTORI,Eric Koiti Okiyama DAL’SASSO,Thaís Carolina da Silva BHERING,Leonardo Lopes OLIVEIRA,Luiz Orlando de |
| author_sort |
MANGARAVITE,Érica |
| title |
A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment |
| title_short |
A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment |
| title_full |
A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment |
| title_fullStr |
A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment |
| title_full_unstemmed |
A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment |
| title_sort |
feasible method to extract dna from the cambium of high-canopy trees: from harvest to assessment |
| description |
ABSTRACT Many tropical trees have high canopies and their leaves are not accessible. Thus, the use of tissue from a more accessible organ (cambium) for DNA extraction may be an alternative for molecular studies. We adapted a feasible methodology for extracting genomic DNA from cambium tissue harvested in the field for the assessment with PCR. We tested three storage conditions (two buffers and a silica gel) and four periods of time after harvest. We used previously described protocols and tested them on three species that occur in Amazonian forests and other biomes: Anadenanthera peregrina var. peregrina, Cedrela fissilis, and Ceiba speciosa. Our protocol obtained suitable PCR-grade genomic DNA for DNA sequencing and microsatellite genotyping. We recommend the use of silica for long-term storage and the buffer with ascorbic acid for short-term storage. |
| publisher |
Instituto Nacional de Pesquisas da Amazônia |
| publishDate |
2020 |
| url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0044-59672020000400335 |
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