Evaluation of the RNA extraction methods in different Ginkgo biloba L. tissues

被引:2
|
作者
Zhang, Xiao-lin [1 ,2 ]
Huang, Xiao-li [1 ]
Li, Jia [1 ]
Mei, Mei [3 ]
Zeng, Wan-qi [1 ]
Lu, Xiu-jun [1 ]
机构
[1] Shenyang Agr Univ, Dept Forestry, 120 Donglin Rd, Shenyang 110866, Liaoning, Peoples R China
[2] Chinese Acad Forestry, Res Inst Forestry, State Key Lab Tree Genet & Breeding, Beijing 100091, Peoples R China
[3] Shenyang Agr Univ, Dept Hort, Shenyang 110866, Peoples R China
基金
国家教育部博士点专项基金资助; 中国国家自然科学基金;
关键词
Ginkgo biloba L; RNA extraction; Quantitative real‐ time PCR; CTAB; Tissues; HIGH-QUALITY RNA; GIBBERELLIN BIOSYNTHESIS; TRANSCRIPTOME ANALYSIS; SEED-GERMINATION; ACID; IMBIBITION; EXPRESSION; GENE; POLYPHENOLS; RESPONSES;
D O I
10.1007/s11756-021-00766-1
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Understanding the essential processes of heredity, mechanisms of metabolism, and evolutionary traits of this ginkgo (Ginkgo biloba L.) in terms of molecular biology is necessary. The extraction of high-quality and high-integrity RNA from ginkgo is one of the basic techniques of molecular biology. Therefore, this study aimed to compare the efficacy of the six commonly used RNA extraction methods for detection. TRIzol, modified TRIzol, cetyltrimethylammonium bromide (CTAB), modified CTAB, sodium dodecyl sulfate (SDS), and RNeasy Plant Mini Kit (Qiagen) methods, were evaluated for their capacity to extract high-quality and high-quantity RNA from the embryo and endosperm of ginkgo. The advantages and disadvantages of each method were analyzed to evaluate the isolation process. The optimized modified CTAB method for embryo prevented the precipitation of RNA for a long period and reduced complex separation and precipitation processes. In addition, the modified TRIzol method is more effective in endosperm. Obvious differences in RNA quality and quantity were observed between the embryo and endosperm when the same method is used. Modified TRIzol, modified CTAB, SDS and RNeasy Plant Mini Kit (Qiagen) methods were applied to RNA extraction from other different ginkgo tissues. The modified CTAB is efficient and recommended for the isolation of good-quality total RNA from all tissues with reduced chemical use, costs, and RNA loss. The results of quantitative real-time PCR confirmed that modified CTAB methods could obtain showing low Ct values and high amplification efficiency and enabled the successful isolation of RNA from all tissues.
引用
收藏
页码:2393 / 2402
页数:10
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