An Optimized DNA Extraction Protocol For Isolation Of High Quality Genomic DNA From Camphor Containing Timber Tree Species Dryobalanops Beccarii DyerJournal: International Journal of Scientific & Technology Research (Vol.5, No. 9)
Publication Date: 2016-09-15
Authors : Wei-Seng Ho; Kit-Siong Liew; Shek-Ling Pang;
Page : 220-223
Keywords : Genomic DNA; CTAB; Drynobanalops beccarii; Camphor; Secondary metabolites; PCR;
Isolation of high-quality genomic DNA from Dryobalanops beccarii is obviously difficult due to the existence of large amounts of camphor and other secondary metabolites. These contaminants will co-precipitate with DNA during DNA isolation and purification processes and therefore resulting in a brownish DNA pellet that is unsuitable for downstream applications. Many DNA isolation protocols are available for various plant tissues however these protocols are inefficient in yielding high-quality amplifiable genomic DNA especially from camphor containing timber tree species. A CTAB based protocol has been optimized for isolating genomic DNA from camphor containing timber tree species. Key steps include 1 using 1 26946-mercaptoethanol and 2 PVP 40 Mr 40000 in the extraction buffer 2 sample incubation time 40 minutes at 65C and 3 DNA precipitation at room temperature 25C. The isolated DNA pellet was transparent colour and the purified genomic DNA is suitable for PCR amplification.
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