Rayani Azadeh, Dehghan Nayeri Fatemeh
Agricultural Biotechnology Department, Faculty of Engineering and Technology, Imam Khomeini International University, Qazvin, Iran.
Biotechnol Lett. 2015 Apr;37(4):927-33. doi: 10.1007/s10529-014-1752-6. Epub 2014 Dec 23.
Seeds of oilseed plants that contain large amounts of oil, polysaccharides, proteins and polyphenols are not amenable to conventional RNA isolation protocols. The presence of these substances affects the quality and quantity of isolated nucleic acids. Here, a rapid and efficient RNA isolation protocol that, in contrast to other methods tested, allows high purify, integrity and yield of total RNA from seeds of sesame, corn, sunflower, flax and rapeseed was developed. The average yields of total RNA from 70 mg oil seeds ranged from 84 to 310 µg with A260/A280 between 1.9 and 2.08. The RNA isolated with this protocol was verified to be suitable for PCR, quantitative real-time PCR, semi-quantitative RT-PCR, cDNA synthesis and expression analysis.
含有大量油脂、多糖、蛋白质和多酚的油料作物种子不适合常规的RNA提取方法。这些物质的存在会影响分离核酸的质量和数量。在此，开发了一种快速高效的RNA提取方法，与其他测试方法相比，该方法能从芝麻、玉米、向日葵、亚麻和油菜籽种子中获得高纯度、完整性好且产量高的总RNA。70毫克油籽的总RNA平均产量在84至310微克之间，A260/A280在1.9至2.08之间。用该方法分离的RNA经证实适用于PCR、定量实时PCR、半定量RT-PCR、cDNA合成和表达分析。
