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以刺山柑为试材,研究比较了改良SDS提取法、RNApure Plant Plus Reagent法、RNAplant Plus Reagent法和RNAprep Pure Plant Kit法提取总RNA的效果,以期寻找一种适于野生刺山柑叶片高质量总RNA最佳提取方法。结果表明:4种方法提取的总RNA均可见28S和18S2条电泳谱带,RNAprep Pure Plant Kit法获得的RNA图谱条带清晰、明亮、稳定,28S rRNA亮度约是18SrRNA的2倍,OD260/OD280为2.00,产率为175.3μg/g,除RNAprep Pure Plant Kit法能够从叶片中提取到完整性好、纯度高及产率高的总RNA外,其它3种方法提取的总RNA均存在降解及DNA和蛋白等污染,经cDNA-SRAP分析检测,此方法获得的RNA反转录后能扩增出清晰稳定的多态性条带,能够满足后续试验的要求。进一步采用RNAprep Pure Plant Kit法从刺山柑幼苗整株、根、茎及叶中能够获得高质量的总RNA,因此,RNAprep Pure Plant Kit法适合刺山柑总RNA的提取。
In this paper, we compared the effects of improved SDS extraction method, RNApure Plant Plus Reagent method, RNAplant Plus Reagent method and RNAprep Pure Plant Kit method on total RNA extraction, in order to find a suitable for wild capers The best quality total RNA extraction method. The results showed that 28S and 18S2 bands were obtained by the 4 methods. The band of RNA obtained by RNAprep Pure Plant Kit was clear, bright and stable. The brightness of 28S rRNA was about 2 times that of 18S rRNA and the OD260 / OD280 Was 2.00, and the yield was 175.3μg / g. The total RNA extracted by the other three methods was degraded except for the RNAprep Pure Plant Kit method which could extract total RNA from the leaves with good integrity, high purity and high yield DNA and protein contamination. The results of cDNA-SRAP analysis showed that this method could amplify clear and stable polymorphic bands after RNA reverse transcription and could meet the requirements of subsequent experiments. Further RNAprop Pure Plant Kit method can obtain high quality total RNA from the whole plant, root, stem and leaf of capparis seedlings. Therefore, RNAprep Pure Plant Kit method is suitable for total RNA extraction of cappi.