An optimized method to\xa0obtain high-quality RNA from different tissues in Lilium davidii var. unicolor

Chunlei Wang,Nana Qi,Changxia Li,Xuemei Hou,Yanyan Luo,Yihua Li,Dongliang Hu,Weibiao Liao

doi:10.1038/s41598-022-06810-7

Abstract

The high quality, yield and purity total RNA samples are essential for molecular experiments. However, harvesting high quality RNA in Lilium davidii var. unicolor is a great challenge due to its polysaccharides, polyphenols and other secondary metabolites. In this study, different RNA extraction methods, namely TRIzol method, the modified TRIzol method, Kit method and cetyltrimethylammonium bromide (CTAB) method were employed to obtain total RNA from different tissues in L. davidii var. unicolor. A Nano drop spectrophotometer and 1% agarose gel electrophoresis were used to detect the RNA quality and integrity. Compared with TRIzol, Kit and CTAB methods, the modified TRIzol method obtained higher RNA concentrations from different tissues and the A260/A280 ratios of RNA samples were ranged from 1.97 to 2.27. Thus, the modified TRIzol method was shown to be the most effective RNA extraction protocol in acquiring RNA with high concentrations. Furthermore, the RNA samples isolated by the modified TRIzol and Kit methods were intact, whereas different degrees of degradation happened within RNA samples isolated by the TRIzol and CTAB methods. In addition, the modified TRIzol method could also isolate high-quality RNA from other edible lily bulbs. Taken together, the modified TRIzol method is an efficient method for total RNA isolation from L. davidii var. unicolor.

Highlights

The high quality, yield and purity total RNA samples are essential for molecular experiments
In Lilium root, RNA samples obtained from the modified TRIzol method showed that the A260/A230 and A260/A280 ratios were closed to 1.5 and 2.1, and RNA samples gained from the cetyltrimethylammonium bromide (CTAB) method showed higher ratios (Table 1)
When RNA was extracted from Lilium leaf using the Kit and CTAB methods, the spectrophotometric results showed that the A260/A230 ratios of RNA ranged from 0.91 to 1.07, and the A260/A280 ratios ranged from 1.69 to 2.01, indicating that the RNA samples were contaminated by proteins, polyphenols and polysaccharides

Summary

Introduction

The high quality, yield and purity total RNA samples are essential for molecular experiments. Different RNA extraction methods, namely TRIzol method, the modified TRIzol method, Kit method and cetyltrimethylammonium bromide (CTAB) method were employed to obtain total RNA from different tissues in L. davidii var. The modified TRIzol method could isolate high-quality RNA from other edible lily bulbs. Kit is used to extract high quality and integrated RNA, which is depending on plant species, genotype and tissue type, and related to the content of polyphenols, polysaccharides and p roteins[10,11]. The quality and yield of RNA obtained by TRIzol method and the modified TRIzol method were compared with each other within different tissues of L. davidii var. We concluded that the modified TRIzol method could be applicable to gain high quality and quantity total RNA in different tissues of L. davidii var. We concluded that the modified TRIzol method could be applicable to gain high quality and quantity total RNA in different tissues of L. davidii var. unicolor containing polyphenols, polysaccharides, proteins and lipids

Methods

Results

Discussion

Conclusion