四种提取青稞种子DNA方法的比较研究OA

To verify the applicability of a self-developed maize single-seed endosperm DNA extraction method that preserves seed viability(referred to as the"patented method")in barley seeds,this study used different barley varieties as materials.DNA was extracted from the seeds by the traditional CTAB method,the modified CTAB method,the alkaline boiling method,and the patented method,respectively.The concentration and purity of the obtained DNA were detected by agarose gel electrophoresis and ultraviolet spectrophotometry.Furthermore,wiht the extracted DNA as a template,conventional PCR,multiplex PCR,and real-time fluorescent quantitative PCR were performed to amplify five genes related to starch synthesis,drought resistance,and disease resistance.The results showed that the DNA extracted from barley seeds by the patented method had a high concentration and good purity and exhibited excellent amplification efficiency in the target gene amplifications,fully meeting the requirements for subsequent molecular biology experiments.Meanwhile,seeds that had undergone partial endosperm excision could still germinate and grow normally and could be used for subsequent related research.This study expands the application range of the patented method to other crop types and provides a high-quality single-seed endosperm DNA extraction method that preserves seed viability for molecular biology research in barley.