Abstract
Isolating and purifying DNA from plants, particularly from species like <i>Cichorium intybus </i>(chicory), poses significant challenges due to the presence of rigid cell walls and high levels of polyphenols and polysaccharides. These compounds can severely hinder the efficiency of traditional DNA extraction protocols, often resulting in poor-quality DNA. Conventional methods, such as the Cetyl-trimethyl-ammonium bromide (CTAB) protocol, are often found inadequate due to these biochemical barriers. In this study, we formulated an optimized DNA isolation protocol for chicory that eliminates the need of liquid nitrogen and phenol during processing. The refined approach resulted in the extraction of high-quality DNA suitable for downstream applications. The effectiveness of the optimized method was validated by polymerase chain reaction (PCR) amplification of the sucrose: sucrose 1-fructosyl transferase (<i>1-SST)</i> gene, a key gene involved in the biosynthesis of inulin- a compound with notable medicinal and nutritional value. The PCR amplification yielded the expected 2.01 kb product, confirming the high quality and integrity of the extracted DNA. Overall, the quantity and quality of the DNA were well-suited for molecular analysis, demonstrating the success of our method. This optimized protocol provides a valuable tool for molecular research on chicory and other plants with similar biochemical challenges, facilitating more efficient genetic studies and holding potential for advancing biotechnological applications.
Published Version
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