A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment

• Autor(a) principal: MANGARAVITE,Érica
• Data de Publicação: 2020
• Outros Autores: TERRA,Vanessa, HATTORI,Eric Koiti Okiyama, DAL’SASSO,Thaís Carolina da Silva, BHERING,Leonardo Lopes, OLIVEIRA,Luiz Orlando de
• Tipo de documento: Relatório
• Idioma: eng
• Título da fonte: Acta Amazonica
• Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0044-59672020000400335
• topic: 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.