摘要
在CTAB法的基础上,经过多方面改进,提取了“红阳”猕猴桃7种组织(果实、雄花、雌花、叶片、种子、皮、根)的总RNA。利用琼脂糖凝胶电泳和超微量核酸蛋白测定仪检测了该方法和试剂盒法提取的RNA的完整性、纯度和浓度。结果显示:采用这两种方法提取到的猕猴桃不同组织的总RNA,能检测到18S和28S两条清晰完整的条带,A 260/A 280值在1.8~2.0间,A 260/A 230值大于2.0。进一步的RT-PCR验证结果表明采用这两种方法提取到的RNA质量较高,均达到了分子生物学实验的要求。因此,本文改良的CTAB法适用于猕猴桃各组织高质量总RNA的大量提取。
Total RNA was successfully isolated by modified CTAB method from seven tissues(fruits,male flowers,female flowers,leaves,seeds,barks,roots)of kiwifruit.The integrity,concentration and purity of total RNA were detected by the agarose gel electrophoresis and nucleic acid/protein assay.The results showed that the total RNA could be observed with two distinct electrophoresis bands of 28S and 18S rRNA,respectively with A 260/A 280 between 1.8 to 2.0 and A 260/A 230 higher than 2.0.RT-PCR analysis also showed that the extracted total RNA by two methods were of good quality,and could be used to other molecular biology experiments.These results indicated this modified CTAB method is suitable for isolating large-scale and high-quality total RNA from a variety of Kiwifruit tissues.
作者
刘林娅
杨那
代玥
罗彩
陈婷
黄亚成
LIU Lin-ya;YANG Na;DAI Yue;LUO Cai;CHEN Ting;HUANG Ya-cheng(School of Biological Science and Technology,Liupanshui Normal University,Liupanshui 553000,China)
出处
《江西农业学报》
CAS
2020年第9期30-34,共5页
Acta Agriculturae Jiangxi
基金
贵州省科技计划项目(黔科合基础[2019]1445号)
贵州省普通高等学校科技拔尖人才项目(黔教合KY字[2018]067)
贵州省普通高等学校特色重点实验室(黔教合KY字[2017]012号)
六盘水师范学院高层次人才科研启动基金项目(LPSSYKYJJ201705、LPSSYKYJJ201706)。
关键词
猕猴桃
多糖多酚
总RNA
提取方法
Kiwifruit
Polysaccharides and polyphenols
Total RNA
Extraction method
