Extraction of Genomic DNA from Different Plant Tissues through Phenol-chloroform Method

Abstract

Background: DNA extraction is a fundamental technique in molecular biology that involves isolating DNA from biological samples for various downstream applications, including PCR and sequencing. Phenol-chloroform DNA extraction is an alternate method of the CTAB method for genomic DNA extraction. This method typically yields more DNA than the CTAB with high purity. It is useful for high molecular weight/long fragment DNA determinations. This study aims to extract DNA from plant tissues using the phenol-chloroform method and compare the response of different plant leaves to this method. Methods: In this study, we provide a step-by-step guide to DNA extraction from five different plant species, including Ficus capensis (Cape fig or Cape banyan),  Ficus exaspirata (Forest Sandpaper), Mangifera indica (Mango),  Gmelina arborea (Gamhar), and  Bauhinia purpurea (Butterfly tree). The DNA isolation protocol involved several steps, such as grinding the plant tissue, adding NaCl solution, using a chloroform-isoamyl alcohol mixture to extract DNA, and further purification with phenol-chloroform-isoamyl alcohol. Isopropanol was used to precipitate the DNA, and ethanol was used to wash the DNA pellet to remove any remaining contaminants. Finally, the DNA pellet was dissolved in TE buffer for storage. Results: The results showed the successful implementation of the phenol chloroform method for isolating DNA from selected plant tissues. Overall, this study provides a comprehensive protocol for DNA extraction from plant tissue using readily available laboratory reagents and equipment. Conclusion: The protocol can be modified to accommodate different plant species and sample types and can facilitate further research in plant genetics and biotechnology.