Resumos do 49° Congresso Brasileiro de Genética
Águas de Lindóia, SP, 16 a 19 de Setembro de 2003
www.sbg.org.br
EXTRACTION OF POLYSACCHARIDE- FREE DNA FROM SHOOT TISSUES OF Cereus
peruvianus Mill. (CACTACEAE) FOR RAPD ANALYSIS
Mangolin, Claudete Aparecida; Silva, Gislaine A Rodrigues; Machado, Maria de Fátima PS
Department of Cell Biology and Genetics, State University of Maringá 87020- 900 Maringá, Paraná, Brazil
[email protected]
Palavras chave: Cereus peruvianus, polysaccharide-free DNA, extraction DNA
The optimization of a method for obtaining quality genomic DNA preparation from the shoot of
Cereus peruvianus adult plant somaclones and from the young shoot of F1 descendents plants of
somaclones (RF1) is provided. These may be consistently amplified via PCR reaction and later
used for RAPD analysis. Six different protocols for plants containing high amounts of polyphenols
and polysaccharides were tested at least 3 times each using four different adult plants and one
young plant per experiment. DNA was obtained from shoot of adult plants by two modified
protocols. While modified Porebiski method consisted in overnight incubation at –20°C instead of 80 °C, prior to last centrifugation, modification in Aljanabi protocol amounted to liquid nitrogen for
grinding of sample and in the overnight incubation at –20°C instead of 60min after the precipitation
with isopropanol and NaCl. DNA from young shoot tissue (RF1 plants) was obtained with five out of
the six tested protocols. DNA extraction from shoot tissues of C. peruvianus analysis indicated that
the requirements and procedures for yield polysaccharide-free DNA varied with type of tissue from
the same specie. Although several authors have researched a relatively simple protocol for DNA
extraction from cactus tissue, tissue-specific responses indicate that empirical testing of di fferent
protocols is indispensable for further analysis of different types and age tissues of cactus species.
Suporte Financeiro: UEM
610