摘要
为了获得能满足高通量测序要求的槟榔叶片总RNA,采用通用植物总RNA提取试剂盒和异硫氰酸胍-苯酚法提取槟榔叶片的总RNA。结果表明:异硫氰酸胍-苯酚法提取的总RNA完整性好、纯度高,通用植物总RNA提取试剂盒不能从槟榔叶片中提取完整的总RNA;70℃,10 min的热处理会导致槟榔总RNA的降解,苯酚/氯仿抽提能得到高纯度、完整性好的总RNA;总RNA质量浓度达到979 mg·L-1,OD260/OD280=1.98,OD260/OD230=2.31,能满足转录组高通量测序的要求。
Total RNA of leaves of Areca nut (Areca catechu L) were extracted by using universal plant total RNA extraction kit and the guanidine isothiocyanate-phenol method to develop an optimum total RNA extraction protocol for Areca leaves for high-throughput sequencing. A better quality of Areca leaf total RNA was extracted by the guanidine isothiocyanate- phenol than the universal plant total RNA extraction kit. Furthermore, two methods, heat treatment and phenol-chloroform extraction, were tried to inactivate DNAase I after total RNA was digested by this enzyme. It was found that the total leaf RNA was degraded when heat treated at 70℃ for 10min, whereas high-purity and high concentration of total leaf RNA was obtained through phenol-chloroform extraction. The quality of total RNA with 0D26o/0D280 = 1.98, 0D260/OD230 = 2.31 and concentration more than 979 mg·L-1, met the standard requirements for high-throughput RNA sequencing.
出处
《热带生物学报》
2015年第1期87-90,共4页
Journal of Tropical Biology
基金
教育部热带作物新品种选育工程中心与作物学重点学科联合资助项目(lhxm-2012-3)
海南大学中西部计划学科重点领域建设项目(ZXBJH-XK001)
关键词
槟榔
RNA提取
高通量测序
异硫氰酸胍-苯酚法
Areca catechu L
RNA extraction
high-throughput sequencing
guanidine isothiocyanate-phenolmethod