College of Horticulture, Nanjing Agricultural University, Weigang, Nanjing 210095, Jiangsu, People's Republic of China.
Mol Biotechnol. 2012 Mar;50(3):229-36. doi: 10.1007/s12033-011-9433-3.
RNA extraction is the first step in the study of gene isolation and expression. However, it is difficult to extract high quantity and quality RNA from tissues containing large quantities of polysaccharides and polyphenols. Peach (Prunus persica), in addition to containing high levels of polysaccharides and polyphenols, is a challenging starting material for RNA isolation using a single method because of different amounts of those substances in diverse tissues. Based on three reported methods, we developed a modified RNA isolation protocol to solve this problem, leading to high quality and quantity of total RNA from peach mesocarp tissues of fruits which were sampled from all developmental stages and different storage periods, as well as from other tissues including flowers, leaves, stems, and roots. With our modified method, 28-650 μg of total RNA was routinely obtained from per gram of fresh material, gave at least a 1.16-fold improvement by compared with those isolated by other seven methods. The RNA extracts were successfully used in downstream applications such as RT-PCR, RACE, and real-time PCR.
RNA 提取是基因分离和表达研究的第一步。然而，从含有大量多糖和多酚的组织中提取高数量和高质量的 RNA 是困难的。桃 (Prunus persica) 除了含有高水平的多糖和多酚外，由于不同组织中这些物质的含量不同，使用单一方法进行 RNA 分离也是一种具有挑战性的起始材料。基于三种已报道的方法，我们开发了一种改良的 RNA 分离方案来解决这个问题，从而从不同发育阶段和不同储存期的果实的桃果肉组织以及其他组织（包括花、叶、茎和根）中获得高质量和高数量的总 RNA。使用我们的改良方法，从每克新鲜材料中通常可获得 28-650μg 的总 RNA，与其他七种方法分离的 RNA 相比，至少提高了 1.16 倍。RNA 提取物成功地用于下游应用，如 RT-PCR、RACE 和实时 PCR。
