https://jopr.mpob.gov.my Thu, 18 Nov 2021 01:47:44 +0000 en-US hourly 1 https://wordpress.org/?v=7.0 https://jopr.mpob.gov.my/isolation-of-high-quality-total-rna-from-various-tissues-of-oil-palm-elaeis-guineensis-for-reverse-transcription-quantitative-real-time-pcr-rt-qpcr/ Thu, 18 Nov 2021 01:33:46 +0000 https://jopr.mpob.gov.my/?p=11534 The extraction of high quality total ribonucleic acid (RNA) has become an essential step for downstream plant molecular biology research. In oil palm, several total RNA extraction protocols have been reported across specific tissues using either conventional method or commercial kits. To our knowledge, there is no specific total RNA extraction method described for oil palm whole fruit which contains lipids, polysaccharides, protein, polyphenols and other secondary metabolites. Here, a modified cetyltrimethylammonium bromidelithium chloride (CTAB-LiCl) method with addition of phenol was established to extract total RNA across various oil palm tissues, especially for whole fruit at eight weeks after anthesis (WAA) and 15 WAA. To assess the extracted total RNA, quality and quantity as well as integrity were evaluated using spectrophotometer and bioanalyser, respectively. It was found that modified CTAB-LiCl method with addition of phenol was able to produce good quality (both A260/A280 and A260/A230 ratios > 1.8) and intact (RIN > 7.4) total RNA. Furthermore, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was successfully performed across all total RNA extracted using two reference genes (PD00569 and pOPEA01332) and one gene of interest, ethylene-responsive transcription factor-3 like, designated as PD00088. The results indicated that the method is suitable for extraction of total RNA from whole fruit as well as other oil palm tissues and is amenable to RT-qPCR.

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