Isolation of High Quality RNA from Canarium odontophyllum Miq. (dabai) Fruits

• Main Author: Lau, Henry Hieng Kui
• Format: Final Year Project Report
• Language: English
• Published: Universiti Malaysia Sarawak, (UNIMAS) 2008
• Subjects: S Agriculture (General); SB Plant culture
• Online Access: http://ir.unimas.my/id/eprint/46560/3/HENRY%20LAU%20%28FULLTEXT%29.pdf; http://ir.unimas.my/id/eprint/46560/

RNA isolation is often the first and most critical step in performing many fundamental molecular biology experiments. Isolation of high quality RNA from Conarium odontophyllum Miq. (dabai) fruits is difficult due to the presence of polysaccharides and substances that bind or co-precipitate with RNA. An efficient and effective protocol is described for isolating high quality RNA from dabai fruits. The described method is a CTAB-based extraction method combined with additional purification steps using butanol and ionic detergent CTAB. The RNA obtained is of high quality and suitable for RT-PCR. The modified RNA isolation protocol, yields RNA of higher purity compared to the original protocol besides having longer purification steps. Butanol is useful in purifying RNA because it helps to remove polysaccharides and proteins that bind to RNA. The optical density indicates that the RNA is relatively pure and suitable for downstream applications and analysis.