摘要
以玫瑰盛开期花瓣为材料,采用RNAiso Plus植物RNA提取试剂盒和改良CTAB法提取总RNA,通过琼脂糖凝胶电泳和紫外分光光度计检测并比较所提取总RNA样品的质量。结果表明:改良CTAB法提取的总RNA较完整,条带清晰,D260nm/D280nm为2.05,D260nm/D230nm为2.12,几乎无降解,纯度较好,质量较高。再采用改良CTAB法提取玫瑰衰老期花瓣及盛开期花器官不同部位组织的总RNA,琼脂糖凝胶电泳、紫外分光光度计检测,并以所提取的玫瑰花组织总RNA为模板进行RT-PCR验证,发现所获得的RNA纯度较好,质量较高,PCR扩增得到的条带较清晰,与目的片段大小一致,完全可以满足进一步分子生物学研究的需要,说明所建立的改良CTAB法是玫瑰花组织总RNA提取的理想方法。
ABSTRACT： Total RNA was isolated from full-opened petals of Rosa rugosa through RNAiso Plus plant RNA extraction kit and improved CTAB method, and the quality of the total RNA was tested by gel electrophoresis and Ultraviolet spec- trophotometer. The results showed that the total RNA extracted by improved CTAB method had clear bands, the value of D260 nm/D280nm and D260 nm/D230 nm was 2. 05 and 2.12 respectively, which indicated that the total RNA scarcely degraded, and its purity and quantity were high. Then, the total RNA in tissues of sentence-stage petals and different organs of full bloom flowers were extracted by improved CTAB method, and the quality of the RNA were determined by gel electropho- resis, Ultraviolet spectrophotometer and RT-PCR. The results showed that the RNA had higher purity and quality, and the PCR product had a distinct band with the same size as theoretical fragment and could satisfy the demands of molecular biology experiment. So the improved CTAB was an appropriate method for extracting the total RNA from Rosa rugosa flower tissues.
出处
《扬州大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2013年第4期104-107,共4页
Journal of Yangzhou University：Agricultural and Life Science Edition
基金
国家自然科学基金资助项目(31000319
31370696)
南通市农业科技创新与产业化项目(HL2012021)
