摘要
为获得高质量的大蒜花器官RNA以用于后续花发育相关基因的研究,以大蒜中多糖多酚含量高的鳞茎为初步实验材料,采用2种试剂盒方法、CTAB法、Trizol法以及改进酸性异硫氰酸胍法提取总RNA,并对提取效果进行比较,筛选出能够得到高质量RNA的天根试剂盒方法和改进酸性异硫氰酸胍法.将改进酸性异硫氰酸胍法进一步用于提取大蒜花器官总RNA,相较于试剂盒方法,获得了质量更好、收率更高的RNA.对提取的大蒜花器官RNA进行完全测序,得到的clean reads占总raw reads比例大于98%,能够满足转录组测序分析的要求.
In order to obtain high-quality RNA from floral organs in garlic for investigating genes related to flower development,five RNA extraction methods including two kits,CTAB,Trizol and improved acidic guanidinium isothiocyanate method were compared using garlic bulbs with high concentration of polysaccharides and polyphenols.The results showed that high-quality RNA could be extracted by Tiangen kit and improved acidic guanidinium isothiocyanate method.The improved acidic guanidinium isothiocyanate method was employed to extract total RNA from floral organs.Compared with kit method,the improved method obtained RNA with better quality and higher yield.The total RNA sequencing was conducted,the result of which showed that the ratio of clean reads to raw reads was above 98%,i.e.the RNA can meet the requirement of transcriptome sequencing analysis.
作者
陈雅楠
尚云涛
范宝莉
刘晓颖
曹宝鑫
王振英
CHEN Ya′nan;SHANG Yuntao;FAN Baoli;LIU Xiaoying;CAO Baoxin;WANG Zhenying(College of Life Sciences,Tianjin Normal University,Tianjin 300387,China;Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin 300387,China;Tianjin Key Laboratory of Water Resources and Water Environment,Tianjin Normal University,Tianjin 300387,China)
出处
《天津师范大学学报（自然科学版）》
CAS
北大核心
2020年第1期34-39,共6页
Journal of Tianjin Normal University：Natural Science Edition
基金
天津市企业科技特派员资助项目(18JCTPJC59100)
天津师范大学应用开发基金资助项目(043135202-XK1707).
关键词
大蒜
花器官
RNA提取
改进酸性异硫氰酸胍法
转录组测序
garlic
floral organ
RNA extraction
improved acidic guanidinium isothiocyanate method
transcriptome sequencing
