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IJSTR >> Volume 5 - Issue 9, September 2016 Edition



International Journal of Scientific & Technology Research  
International Journal of Scientific & Technology Research

Website: http://www.ijstr.org

ISSN 2277-8616



An Optimized DNA Extraction Protocol For Isolation Of High Quality Genomic DNA From Camphor Containing Timber Tree Species, Dryobalanops Beccarii Dyer

[Full Text]

 

AUTHOR(S)

Wei-Seng Ho, Kit-Siong Liew, Shek-Ling Pang

 

KEYWORDS

Genomic DNA, CTAB, Drynobanalops beccarii, Camphor, Secondary metabolites, PCR

 

ABSTRACT

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% β-mercaptoethanol and 2% PVP 40 (Mr 40,000) 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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