Abstract:
The extraction of high-quality RNA from cotton (Gossypium spp.) is challenging because of the presence of high polyphenolics, polysaccharides, quinones, and other secondary metabolites. A high-throughput RNA extraction protocol is a prerequisite. This Triton-X-100-based RNA extraction method utilizes Polyvinyl pyrrolidone polymer (PVPP) treatment which efficiently removes phenolics, and the application of Lithium chloride (LiCl) has been found that successfully precipitated the high-quality RNA from cotton tissue. Cytoplasmic male sterility (CMS) is a maternally inherited trait associated with specific mitochondrial genome rearrangements or mutations. The suitability of RNA extracted from Cotton CMS lines was assessed. cDNA was synthesized from RNA and assayed for mitochondrial genes (cox3, nad3, nad9) associated with male sterility. This paper discuss the advantages and limitation of this protocol over existing protocol for RNA extraction for polyphenolics-rich plant tissue.
摘要:
从棉花中提取优质RNA(棉属。)是具有挑战性的,因为存在高多酚,多糖,醌,和其他次生代谢产物。高通量RNA提取方案是先决条件。这种基于Triton-X-100的RNA提取方法利用聚乙烯吡咯烷酮聚合物(PVPP)处理,可有效去除酚类,氯化锂(LiCl)的应用已被发现成功地从棉花组织中沉淀出高质量的RNA。细胞质雄性不育(CMS)是与特定线粒体基因组重排或突变相关的母系遗传性状。评估从棉花CMS系提取的RNA的适合性。从RNA合成cDNA,并测定与雄性不育相关的线粒体基因(cox3、nad3、nad9)。本文讨论了该协议相对于富含多酚的植物组织的RNA提取的现有协议的优点和局限性。