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农学学报 ›› 2015, Vol. 5 ›› Issue (10): 93-96.doi: 10.11923/j.issn.2095-4050.cjas15010014

• 农业工程 农业机械 生物技术 食品科学 • 上一篇    下一篇

玫瑰花柱总RNA提取方法的比较研究

崔院院,于晓艳   

  1. 山东农业大学林学院,山东农业大学林学院
  • 收稿日期:2015-01-10 修回日期:2015-03-12 接受日期:2015-03-20 出版日期:2015-10-30 发布日期:2015-10-30
  • 通讯作者: 于晓艳 E-mail:1041233596@qq.com
  • 基金资助:
    国家自然科学基金“玫瑰β-l,3-葡聚糖合成酶基因和β-l,3-葡聚糖酶基因克隆与调控授粉亲和性的功能分析”(31200524)

Comparison of Methods for Total RNA Extraction from Rosa rugosa.

  • Received:2015-01-10 Revised:2015-03-12 Accepted:2015-03-20 Online:2015-10-30 Published:2015-10-30

摘要: 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。

关键词: 大花黄牡丹, 大花黄牡丹, 变异材料, 光合特性, 拉萨

Abstract: 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.

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