An innovative optimized protocol for high-quality genomic DNA extraction from recalcitrant Shea tree (Vitellaria paradoxa, C.F. Gaertn) plant and its suitability for downstream applications.
DNA purity; Downstream applications; Genomic DNA extraction protocol; Polymerase chain reaction; Sanger sequencing; DNA; Polysaccharides; Random Amplified Polymorphic DNA Technique; Trees; Genomics; Molecular Biology; Genetics; General Medicine
Abstract :
[en] [en] BACKGROUND: It is not always easy to find a universal protocol for the extraction of genomic DNA (gDNA) from plants. Extraction of gDNA from plants such as shea with a lot of polysaccharides in their leaves is done in two steps: a first step to remove the polysaccharides and a second step for the extraction of the gDNA. In this work, we designed a protocol for extracting high-quality gDNA from shea tree and demonstrate its suitability for downstream molecular applications.
METHODS: Fifty milligrams of leaf and root tissues were used to test the efficiency of our protocol. The quantity of gDNA was measured with the NanoDrop spectrometer and the quality was checked on agarose gel. Its suitability for use in downstream applications was tested with restriction enzymes, SSRs and RAPD polymerase chain reactions and Sanger sequencing.
RESULTS: The average yield of gDNA was 5.17; 3.96; 2.71 and 2.41 µg for dry leaves, dry roots, fresh leaves and fresh roots respectively per 100 mg of tissue. Variance analysis of the yield showed significant difference between all tissue types. Leaf gDNA quality was better compared to root gDNA at the absorbance ratio A260/280 and A260/230. The minimum amplifiable concentration of leaf gDNA was 1 pg/µl while root gDNA remained amplifiable at 10 pg/µl. Genomic DNA obtained was also suitable for sequencing.
CONCLUSION: This protocol provides an efficient, convenient and cost effective DNA extraction method suitable for use in various vitellaria paradoxa genomic studies.
Silué, Souleymane; Department of Biochemistry-Genetics, Faculty of Biological Sciences, Educational and Research Unit of Genetic, University of Peleforo Gon Coulibaly (UPGC), BP 1328, Korhogo, Côte d'Ivoire
Yao, Saraka Didier Martial; Department of Biochemistry-Genetics, Faculty of Biological Sciences, Educational and Research Unit of Genetic, University of Peleforo Gon Coulibaly (UPGC), BP 1328, Korhogo, Côte d'Ivoire
De Clerck, Caroline ; Université de Liège - ULiège > TERRA Research Centre > Plant Sciences
Shumbe, Léonard ; Université de Liège - ULiège > Département GxABT > Plant Sciences
Diarrassouba, Nafan; Department of Biochemistry-Genetics, Faculty of Biological Sciences, Educational and Research Unit of Genetic, University of Peleforo Gon Coulibaly (UPGC), BP 1328, Korhogo, Côte d'Ivoire
Fofana, Inza Jésus; Department of Biochemistry-Genetics, Faculty of Biological Sciences, Educational and Research Unit of Genetic, University of Peleforo Gon Coulibaly (UPGC), BP 1328, Korhogo, Côte d'Ivoire
Alabi, Taofic ; Université de Liège - ULiège > Département GxABT > Gestion durable des bio-agresseurs ; Department of Biochemistry-Genetics, Faculty of Biological Sciences, Educational and Research Unit of Genetic, University of Peleforo Gon Coulibaly (UPGC), BP 1328, Korhogo, Côte d'Ivoire
An innovative optimized protocol for high-quality genomic DNA extraction from recalcitrant Shea tree (Vitellaria paradoxa, C.F. Gaertn) plant and its suitability for downstream applications.
Publication date :
22 January 2024
Journal title :
Molecular Biology Reports
ISSN :
0301-4851
eISSN :
1573-4978
Publisher :
Springer Science and Business Media B.V., Netherlands
We acknowledge the support of the plant Genetics laboratory at Gembloux Agro Bio-Tech and the Department of Biochemistry-Genetics, Educational and Research Unit of Genetics at the Péléforo Gon Coulibaly University in Korhogo, which made this study possible.
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