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Evaluation of the RNA extraction methods in different Ginkgo biloba L. tissues
- Source :
- Biologia. 76:2393-2402
- Publication Year :
- 2021
- Publisher :
- Springer Science and Business Media LLC, 2021.
-
Abstract
- 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.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Plant Science
01 natural sciences
Biochemistry
Endosperm
03 medical and health sciences
chemistry.chemical_compound
Genetics
Sodium dodecyl sulfate
Molecular Biology
Ecology, Evolution, Behavior and Systematics
Chromatography
biology
Chemistry
Ginkgo biloba
Ginkgo
Extraction (chemistry)
RNA
Cell Biology
biology.organism_classification
030104 developmental biology
Trizol
Animal Science and Zoology
RNA extraction
010606 plant biology & botany
Subjects
Details
- ISSN :
- 13369563 and 00063088
- Volume :
- 76
- Database :
- OpenAIRE
- Journal :
- Biologia
- Accession number :
- edsair.doi...........ef6fd4a82dc66e2cbc06e27fcfc2e494