Optimization of DNA extraction from fresh leaf tissues  of Melanoxylon brauna (Fabaceae).
Melanoxylon brauna (Fabaceae - Caesalpinioideae) is an endemic and  valuable hardwood tree species in the Brazilian Atlantic rainforest; it is  comparable to African ebony wood. We tested three protocols of DNA extraction  based on the citrimonium bromide (CTAB) method and evaluated the quantity,  purity and integrity of the DNA. We also determined whether these procedures  interfere with PCR amplification in order to develop a protocol for M.  brauna. We found that the quality and integrity of DNA were improved with  the use of proteinase K in the extraction buffer and by modifications in the  centrifugation speed. The lowest concentration of DNA was obtained with Doyle  and Doyle's protocol (5.42 ng/μL). Ferreira and Grattapaglia's protocol modified for M.  brauna provided the most DNA (36.89 ng/μL) and the highest quality DNA (purity ratio of 1.80  nm). The original Ferreira and Grattapaglia protocol provided 13.42 ng/μL DNA; however, the purity ratio (1.44 nm)  indicates protein contamination. PCR results showed that Ferreira and  Grattapaglia's protocol modified for M. brauna gave satisfactory  quantity and purity of DNA for molecular studies.
Departamento de Ciências Biológicas, Universidade Estadual do Sudoeste da Bahia, Jequié, BA, Brasil. dani_borges88@hotmail.com
D B
DB
Borges
M B
MB
Amorim
A M
AM
Waldschmidt
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E
Mariano-Neto
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CV
Vivas
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DG
Pereira
D018744:DNA, Plant / Q000235:genetics* / Q000302:isolation & purification*; D007887:Fabaceae / Q000235:genetics*; D018895:Microsatellite Repeats / Q000235:genetics; D018515:Plant Leaves / Q000235:genetics*
D018744:DNA, Plant