适合转录组测序的芒果不同组织RNA试剂盒提取方法研究

Study on kits extraction methods of RNA suitable for transcriptome sequencing from different tissues in Mangifera indica

为了探寻能够满足芒果各组织转录组测序要求的高质量RNA提取方法,以‘桂热芒82号’的叶片、花和果实为材料,从RNA提取质量和产率、琼脂糖电泳和RT-PCR结果分析、RNA测序质量评价这3个方面,对常用的3种试剂盒的提取效果进行了实验研究和评价分析。结果表明:以来自于天根生化科技有限公司的DP 419试剂盒提取的芒果各组织的RNA质量均较好,提取的RNA产率远高于其余两种试剂盒的提取产率;除以美伯生物医药技术有限公司的RM 103试剂盒提取芒果各组织所得RNA,经琼脂糖电泳未见有5S条带出现外,以其余两种试剂盒提取的芒果各组织的RNA都存在28S、18S、5S条带且RT-PCR分析结果理想;对以3种试剂盒提取所得的RNA进行完全测序,结果表明,3种试剂盒提取的RNA完全测序都大于98.99%,能满足RNA-Seq分析的要求。文中综合上述评判因素分析认为,天根生化科技有限公司的DP 419试剂盒是适用于芒果叶片、花和果实RNA提取的最优试剂盒。

In order to study extraction methods of high-quality RNAs from each tissue in Mangifera indica,which could meet the requirement of transcriptome sequencing,leaves,flowers and fruits of‘Guiremang No.82’were used as materials,extraction effects of three common kits were researched and evaluated,from three aspects of RNA extraction quality and yield,agarose electrophoresis and RT-PCR results analysis,and RNA sequencing quality evaluation.The results showed that extracted RNAs from all tissues in M.indica had good quality by using the DP419 kit from Tiangen Biochemical Technology Co.,Ltd.,and extracted RNA yield was much higher than those by using the other two kits.Extracted RNAs from each tissue in M.indica showed no 5S band through agarose electrophoresis by using the RM103 kit from Meibo Biomedical Technology Co.,Ltd..Those by using the other two kits had 28S,18S and 5S bands,and their RT-PCR results were ideal.The results of showed that total RNA sequencing extracted from the three kits was more than 98.99%,and could meet the requirement of RNA-seq analysis.In conclusion,the DP419 kit from Tiangen Biochemical Technology Co.,Ltd.was the optimal one for extracting RNAs from leaves,flowers and fruits in M.indica.