An Optimized DNA Extraction Protocol For Isolation Of High Quality Genomic DNA From Camphor Containing Timber Tree Species, Dryobalanops Beccarii Dyer
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 bro...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | E-Article |
| Language: | English |
| Published: |
IJSTR
2016
|
| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/14331/1/An%20Optimized%20DNA%20Extraction%20Protocol%20For%20Isolation%20Of%20High%20Quality%20%28abstract%29.pdf http://ir.unimas.my/id/eprint/14331/ http://www.ijstr.org/research-paper-publishing.php?month=sep2016 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Summary: | 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 65°C, and 3) DNA precipitation at room temperature (25°C). The isolated DNA pellet was transparent colour and the purified genomic DNA is suitable for PCR amplification.
Wei-Seng Ho, Kit-Siong Liew, Shek-Ling Pang |
|---|