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

• Main Author: Liew, Kit Siong
• Format: Final Year Project Report
• Language: English
• Published: Universiti Malaysia Sarawak (UNIMAS) 2007
• Subjects: SB Plant culture
• Online Access: http://ir.unimas.my/id/eprint/7609/3/LIEW%20KIT%20SIONG.pdf; http://ir.unimas.my/id/eprint/7609/
• Institution: Universiti Malaysia Sarawak

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.