CAJ | 학술논문

为获得一种玫瑰(Rosa rugosa)花柱总RNA提取的理想方法，为后续相关基因工程研究奠定基础。以玫瑰花柱为材料，用核酸蛋白仪和凝胶电泳法比较了改良CTAB法、Trizol法以及EASY spin Plus植物RNA提取试剂盒法提取的玫瑰花柱总RNA的纯度、浓度及完整性，利用RT-PCR反应检测了其反转录产物用于基因扩增的有效性。结果表明：改良CTAB法提取的总RNA纯度、浓度较高，但存在严重降解现象；Trizol法提取的总RNA纯度和浓度极低；EASY spin Plus植物RNA提取试剂盒法提取的总RNA纯度和浓度较高，条带清晰，且28S条带亮度是18S条带亮度的2倍，将其反转录获得的cDNA用于RT-PCR，能够扩增出清晰的特异性条带。综上所述：EASY spin Plus植物RNA提取试剂盒法从玫瑰花柱中提取的RNA质量最好，完全能够满足后续的玫瑰分子生物学研究要求。
위획득일충매괴(Rosa rugosa)화주총RNA제취적이상방법，위후속상관기인공정연구전정기출。이매괴화주위재료，용핵산단백의화응효전영법비교료개량CTAB법、Trizol법이급EASY spin Plus식물RNA제취시제합법제취적매괴화주총RNA적순도、농도급완정성，이용RT-PCR반응검측료기반전록산물용우기인확증적유효성。결과표명：개량CTAB법제취적총RNA순도、농도교고，단존재엄중강해현상；Trizol법제취적총RNA순도화농도겁저；EASY spin Plus식물RNA제취시제합법제취적총RNA순도화농도교고，조대청석，차28S조대량도시18S조대량도적2배，장기반전록획득적cDNA용우RT-PCR，능구확증출청석적특이성조대。종상소술：EASY spin Plus식물RNA제취시제합법종매괴화주중제취적RNA질량최호，완전능구만족후속적매괴분자생물학연구요구。
To select the suitable method for total RNA extraction from the style of Rosa rugosa, and provide foundation for related genetic engineering research, total RNA was extracted with three methods, including modified CTAB, Trizol and EASY spin Plus Plant RNA Kit. And the purity, concentration and completeness were compared by nucleic acid protein detector and gel electrophoresis. The effectiveness of gene amplification of reverse transcription product was checked by RT-PCR reaction. The results showed that the purity, concentration and completeness of total RNA extracted by modified CTAB and EASY spin Plus Plant RNA Kit were higher than that by Trizol. But the bands of total RNA extracted by modified CTAB had serious degradation and the Kit was clear. And the 28S was two times as bright as 18S. The reverse transcription of cDNA obtained for RT-PCR could clearly amplify specificity band. These results demonstrate that the RNA isolated by EASY spin Plus Plant RNA Kit is of sufficient quality and can satisfy subsequent molecular study for Rosa rugosa.