Evaluation of DNA Extraction Methods for RAPD, SSR and AFLP Analyses of Wild Rose Species

摘要

The current study aims to evaluate a rapid, simple and robust method of DNA extraction for AFLP analysis of 12 wild rose species (Rosa brunonii, R. cathayensis, R. moschata, R. multiflora, R. wichurriana, R. indica, R. alba, R. macrophylla, R. tomentosa, R. canina, R. damascena, R. bourboniana and the Fl progeny of R. damascena and R. bourboniana). Extraction of quality DNA from wild rose species is difficult as they contain high levels of polysaccharides and polyphenols. Four DNA extraction protocols were compared: two commercial kits from Qiagen and AuPrep, CTAB and a modified CTAB protocol. The protocols were evaluated in terms of yield, purity, restrictability and amplifiability of recovered DNA. The yield and quality of genomic DNA was considerably affected when commercial kits and common CTAB protocol were utilized for DNA isolation. The modified phenol free, CTAB procedure involving a washing step before extraction was the most successful extraction method giving optimum yields (900-1750 ug/g) of quality DNA that was amenable to restriction digestion and polymerase chain reaction (PCR) analyses - RAPD, SSR, AFLP.