适于转录组测序的枣不同器官总RNA提取方法筛选

Screening of total RNA extraction methods for RNA-sequences from different organs in Ziziphus jujuba

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。

In order to obtain the methods for high-quality total RNA extraction from flower, fruit bearing branch and fruit in Ziziphusjujuba, taking Z. jujuba cv. ＇Zhongqiusucui＇ as materials, and the RNA extraction effects were compared using two kinds of total RNA extraction kits （Ambiogen and Autolab） and modified CTAB method based on the Autolab RNA extraction kit. The results showed that OD260/OD280 values of the RNA extracted by using the three RNA extraction methods had no significant differences, but a part of RNAs obtained by the two RNA extraction kits were degraded. The RNA mass concentrations from flower, fruit bearing branch and fruit were 126, 284 and 222 ng/μL by using the RNA extraction kit of Ambiogen; the RNA mass concentrations were 307, 402 and 266 ng/μL by using the RNA extraction kit of Autolab; and the RNA mass concentrations were 401,417 and 296 ng/μL by using the modified CTAB method based on the Autolab RNA extraction kit, respectively. The modified CTAB method could generate high-integrity, high-concentration and high-purity RNA that could meet the request oftranscriptome sequencing, compared with the two extraction kits.