1. [High-quality total RNA extraction from medicinal plant Aquilaria sinensis]
- Author
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Hong-Qing, Wu, Lei, Wang, Wei, Guo, Xiao-Xia, Gao, Ling, Bai, and Wei-Min, Zhang
- Subjects
Plant Leaves ,Plants, Medicinal ,Plant Stems ,RNA, Plant ,Thymelaeaceae ,Spectrophotometry, Ultraviolet ,Guanidines ,Thiocyanates - Abstract
It is quite difficult to isolate total RNA from Aquilaria sinensis due to the complexity of its secondary metabolites. In order to obtain the high-quality RNA, we investigated sampling method and RNA extraction from both normal and agilawood tissues of the plant.Five methods such as modified CTAB-LiCl method, modified CTAB-LiCl method with no water-bath, SDS-acid phenol method, modified guanidinium thiocyanate-CTAB method and EASYspin RN09 RNA isolation kit were used to extract RNA from leaves, branches, white wood samples, agilawood samples of A. sinensis, respectively.Total RNA was stable for 5 hours at room temperature after extracted from A. sinensis branches. EASYspin RN09 RNA isolation kit was the best extraction method for the leaf while modified guanidinium thiocyanate-CTAB method was the best for the other samples.A great quantity of RNAs can be isolated from samples of white wood and agilawood wood, respectively. Their ratios of 28S : 18S are in the range of 1.2 - 1.4 and RINs are between 6.8 - 7.5. These RNAs can be applied in transcriptome sequencing.
- Published
- 2014