Isolation of high quality genomic dna from dryobnanalops beccarii dyer tissues rich in camphor
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,...
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| Format: | Final Year Project Report |
| Language: | English |
| Published: |
Universiti Malaysia Sarawak (UNIMAS)
2007
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| Online Access: | http://ir.unimas.my/id/eprint/7609/3/LIEW%20KIT%20SIONG.pdf http://ir.unimas.my/id/eprint/7609/ |
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| Institution: | Universiti Malaysia Sarawak |
| Language: | English |
| Summary: | 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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