TY - JOUR AU - Kansup, Jeeraporn AU - Kumchoo, Tanavadee AU - Chanroj, Vipavee AU - Ngorian, Suphawadee AU - Amawan, Suwaluk AU - Wongtiem, Prapit PY - 2021/05/05 Y2 - 2024/03/29 TI - A Rapid, Economical and Hazardous Organic Solvent Free Method for DNA Extraction from Cassava JF - Thai Agricultural Research Journal JA - Thai Agric.Res.J. VL - 39 IS - 2 SE - Technical or research paper DO - 10.14456/thaidoa-agres.2021.16 UR - https://li01.tci-thaijo.org/index.php/thaiagriculturalresearch/article/view/248057 SP - AB - <p>The extraction of good quality DNA from leaves of cassava is a complicated process<br>due to the presence of interfering substances such as polyphenols and polysaccharides that<br>affect the quality of DNA. In hexadecyltrimethylammonium bromide or CTAB method which<br>is widelyusedfor DNA extraction,hazardousorganic solvents suchasphenol andchloroform<br>are required to remove these interfering substances. The whole process does take<br>approximately 130 minutes making it unsuitable for extracting large number of DNA samples.<br>The objective of this research was to develop a DNA extraction method from cassava leaves<br>which is rapid, free of harmful organic solvents, does not use liquid nitrogen, as well as cost<br>and time effective. The DNA extraction method developed in this research was based on<br>sodium dodecyl sulfate/sodium chloride or SDS/NaCl method. The SDS/NaCl+PVP method<br>yielded DNA amount in the range of 1,431.4 - 1,510.3 µg per 1g of leaf sample with absorption<br>ratios A260/A280 of 1.74 - 1.90. Agarose gel electrophoresis showed the intact, sharp and clear<br>DNA bands with little degradation. The extracted DNA could be used in restriction analysis<br>and as template in polymerase chain reaction (PCR) using many types of primer including<br>housekeeping gene, universal, SCAR, EST and SSR primers in both of single and multiplex<br>PCR. It indicated that the DNA extracted by SDS/NaCl+PVP method had purity, good quality<br>and was suitable for molecular analyses such as restriction digestion and PCR<br>amplification.The wholeprocessof DNA extractionusing SDS/NaCl+PVP methodtook about<br>48 minutes.</p> ER -