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一种适用于逆转录定量实时PCR(RT-qPCR)的从油棕组织中快速提取RNA的方法。

A rapid RNA extraction method from oil palm tissues suitable for reverse transcription quantitative real-time PCR (RT-qPCR).

作者信息

Badai Siti Suriawati, Rasid Omar Abd, Parveez Ghulam Kadir Ahmad, Masani Mat Yunus Abdul

机构信息

Advanced Biotechnology and Breeding Centre (ABBC) Division, Malaysian Palm Oil Board (MPOB), 6, Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor Malaysia.

出版信息

3 Biotech. 2020 Dec;10(12):530. doi: 10.1007/s13205-020-02514-9. Epub 2020 Nov 12.

Abstract

Cetyltrimethylammonium bromide (CTAB) is the preferred detergent in RNA extraction of oil palm tissues. However, the CTAB-based protocol is time-consuming. In this study, a combination of the CTAB-based method and silica-based purification reduced the extraction time from two days to five hours. Quality of total RNA from 27 different tissues of oil palm was shown to have an RNA integrity number (RIN) value of more than seven. The extracted RNA was evaluated by RT-qPCR using three reference oil palm genes (, , and ) and three putative mesocarp-specific transcripts annotated as WRKY DNA-binding protein 70 () metallothionein () and pentatricopeptide repeat () genes. Tissue-specific expression profiling across complete developmental stages of mesocarp and vegetative tissues was determined in this study. Overall, the RNA extraction protocol described here is rapid, simple and yields good quality RNAs from oil palm tissues.

摘要

十六烷基三甲基溴化铵(CTAB)是油棕组织RNA提取中首选的去污剂。然而,基于CTAB的方案耗时较长。在本研究中,基于CTAB的方法与基于硅胶的纯化相结合,将提取时间从两天缩短至五小时。结果表明,来自油棕27种不同组织的总RNA的RNA完整性数(RIN)值大于7。使用三个油棕参考基因(、和)以及三个注释为WRKY DNA结合蛋白70()、金属硫蛋白()和五肽重复()基因的假定中果皮特异性转录本,通过RT-qPCR对提取的RNA进行评估。本研究确定了中果皮和营养组织完整发育阶段的组织特异性表达谱。总体而言,本文所述的RNA提取方案快速、简单,并且能从油棕组织中获得高质量的RNA。

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