Isolation of high quality genomic dna from dryobnanalops beccarii dyer tissues rich in camphor

Liew, Kit Siong (2007) Isolation of high quality genomic dna from dryobnanalops beccarii dyer tissues rich in camphor. [Final Year Project Report] (Unpublished)

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Abstract

The isolation of high-quality intact genomic DNA is obviously difficult in a variety of plant species due to the presence of high levels of polysaccharides, polyphenolics 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. Most plant genomic DNA isolation protocols for various plant tissues have been established; however these protocols are inefficient in yielding high-quality amplifiable genomic DNA especially from camphor containing timber species, Drynobanalops beccarii Dyer. In this project, the total genomic DNA of Drynobanalops beccarii was successfully isolated using a re-optimized CTAB method based on the Murray & Thompson (1980) and Doyle & Doyle (1990). The modifications include: 1) using 1% =-mercaptoethanol and 2% PVP (Mr 40000) in the extraction buffer, 2) sample incubation time, 40 minutes at 65°C; and 3) DNA precipitation at room temperature (25°C). The isolated DNA pellet was in transparent colour and yielded amplifiable genomic DNA for RAPD-PCR.

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