适于转录组测序的枣不同器官总RNA提取方法筛选

摘要

In order to obtain themethodsfor high-quality total RNA extractionfromlfower,fruit bearing branchandfruit inZiziphus jujuba, takingZ. jujuba cv.'Zhongqiusucui'asmaterials,and the RNA extraction effectswere compared using twokinds of total RNA extractionkits (Ambiogenand Autolab)andmodiifed CTABmethod based on the Autolab RNA extractionkit. Theresults showed thatOD260/OD280values of the RNA extracted by using the three RNA extractionmethods had no signiifcant differences, buta part of RNAs obtained by the two RNA extractionkitswere degraded. The RNAmass concentrationsfromlfower,fruit bearing branchandfruitwere 126, 284and 222 ng/μL by using the RNA extractionkit of Ambiogen; the RNAmass concentrationswere 307, 402and 266 ng/μL by using the RNA extractionkit of Autolab;and the RNAmass concentrationswere 401, 417and 296 ng/μL by using themodiifed CTABmethod based on the Autolab RNA extractionkit,respectively. Themodiifed CTABmethod couldgenerate high-integrity, high-concentrationand high-purity RNA that couldmeet therequest of transcriptome sequencing, comparedwith the two extractionkits.%为获得适用于转录组测序的枣花、结果枝和果实的总RNA提取方法,以中秋酥脆枣为材料,比较分析了Ambiogen和Autolab 2种总RNA提取试剂盒和基于Autolab试剂盒改良的CTAB法的总RNA的提取效果.结果表明:3种方法提取RNA的OD260/OD280差别不大,但用2种试剂盒提取的RNA有降解,其中Ambiogen试剂盒提取的花、结果枝和果实3个器官的RNA质量浓度分别为126、284和222 ng/μL;Autolab试剂盒提取的RNA质量浓度分别为307、402和266 ng/μL;基于Autolab试剂盒改良的CTAB法提取的RNA质量浓度分别为401、417和296 ng/μL.与2种试剂盒相比,基于Autolab试剂盒改良的CTAB法提取出来的RNA完整性好、纯度和浓度高,能够满足转录组测序的要求.