An optimized method to obtain high-quality RNA from different tissues in Lilium davidii var. unicolor

Abstract

Background: The high quality, high yield and purity RNA samples are essential for subsequent molecular experiments such as RT-PCR, qPCR and RNA-seq. However, harvest high quality RNA samples from different tissues in Lilium davidii var. unicolor is a great challenge due to its polysaccharides, polyphenols and other secondary metabolites. In this study, three RNA extraction methods, namely modified TRIzol method, Kit and CTAB methods were reported to obtain the total RNA from Lilium davidii var. unicolor, and the efficient RNA extraction protocol (modified TRIzol method) was described. Results: A Nano drop spectrophotometer and 1% gel electrophoresis were used to detect the RNA quality and integrity. Compared with Kit and CTAB methods, the higher RNA concentrations from different tissues were obtained and the A260/280 ratios of RNA samples were ranged from1.97 to 2.27 when using the modified TRIzol method. None of intact RNA bands were gained from the modified CTAB method, indicating that the RNA samples isolation were degraded. As a result of the modified TRIzol method, the RNA samples in the different tissues from Lilium presented clear 28S and 18S bands.Conclusions: Here, modified TRIzol method is an easy, efficient, and low-cost method for RNA isolation from Lilium davidii var. unicolor. Thus, modified TRIzol method is sufficient to gain eligible RNA to support further molecular experiments in Lilium davidii var. unicolor.

Development and Validation of Rapid Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification for Detection of Rift Valley Fever Virus

“…Tese fndings agree with the fndings of Wang et al [36], in terms of purity. However, the concentration of extracted total RNA was found to be averagely higher compared to that extracted by Wang et al [36]. Te purity and integrity of the extracted total RNA were key to downstream processing [38].…”

“…Te total RNA was extracted using the TRIzol reagent method. Te selection of the TRIzol reagent method was based on previous studies that intimated that the TRIzol reagent method was superior to the current column-based RNA extraction kits in the generation of high-quality RNA [33,35,36].…”

“…Since nucleic acid maximumly absorbs light at A260 nm, the concentration of total RNA extracted was automatically calculated by multiplying the absorbance at A260 nm wavelength with an extinction coefcient factor of 40 [37]. A pure RNA sample must have A260/A280 and A260/A230 ratios at 1.8-2.2 and > 1.7, respectively [36]. During this study, all values of total extracted RNA were within the recommended ratio ranges of 1.91-2.01 and 1.86-2.00 for A260/A280 and A260/A230 ratios, respectively (Table 1).…”