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水茄果实中富含多糖、蛋白、色素和其他代谢物质,影响其总RNA提取的质量。本研究中比较Trizol法、改良Trizol法、改良CTAB法和两种试剂盒法5种方法提取的水茄果实总RNA,用琼脂糖凝胶电泳检测总RNA的完整性,Onedrop光谱仪检测总RNA的浓度和纯度。结果表明:改良Trizol法最为有效,该方法提取的总RNA在琼脂糖凝胶上得到清晰的28S和18S条带,OD_(260nm)/OD280nm值在1.8~2.1之间、OD_(260nm)/OD_(230nm)的比值大于2.0;改良CTAB法获得的RNA质量较好,但步骤较为繁琐,耗时长;Trizol法和试剂盒法得到总RNA浓度较低。RT-PCR试验进一步表明,以改良Trizol法获得的总RNA可用于基因分离,所得条带清晰,与目的片段大小一致且比对正确。
Water eggplant is rich in polysaccharides, proteins, pigments and other metabolites, affecting the quality of its total RNA extraction. In this study, Trizol method, modified Trizol method, modified CTAB method and two kit methods were used to extract total RNA of watery fruits, the integrity of total RNA was detected by agarose gel electrophoresis, the total RNA was detected by Onedrop spectrometer Concentration and purity. The results showed that the modified Trizol method was the most effective method. The total RNA extracted by this method showed clear 28S and 18S bands on the agarose gel. The OD 260nm / OD 280nm ranged from 1.8 to 2.1 and the OD 260nm OD_ (230nm) ratio is greater than 2.0. The quality of RNA obtained by modified CTAB method is better, but the steps are more cumbersome and time-consuming. The total RNA concentration obtained by Trizol method and kit method is lower. RT-PCR test further showed that the total RNA obtained by the modified Trizol method can be used for gene isolation, the resulting bands are clear, consistent with the size of the target fragment and the correct alignment.