摇蚊基因组DNA的提取方法研究

Research on Extraction Method of Genomic DNA from Chironomidae

采用动物基因组DNA快速抽提试剂盒法、SDS法、CTAB法、NaCl法4种方法提取摇蚊基因组DNA.通过紫外分光光度计分别检测所得样品DNA在波长为260nm和280nm的吸光值,根据A260/A280的比值检测其纯度和浓度.采用PCR技术获得摇蚊COI基因,用琼脂糖凝胶电泳检测PCR-COI.综合分光光度法与电泳结果筛选出最适合提取摇蚊基因组DNA的方法.结果表明:SDS法所得样品DNA纯度最高,试剂盒法所得DNA浓度最大.同时综合比较DNA纯度与浓度分析得出:试剂盒法SK8222(改良)提取摇蚊基因组DNA为最佳方法,其次为SDS法或CTAB法(改良).

This research used four methods （ animal genomic DNA rapid extraction kit method , SDS method, CTAB method and NaCl method ） to extract the genomic DNA of Chironomidae , and then re-spectively measured the absorbance value of DNA sample by ultraviolet spectrophotometer in optical wave -length of 260 nm and 280 nm,and checked its purity and concentration according to the ratio of A 260 /A280.Finally, the PCR results of the COI gene of Chironomidae were tested with agarose gel electropho -resis, through the analysis of comprehensive spectrophotometry and electrophoresis results selecting the most suitable method for extracting Chironomidae genomic DNA .The results show that we can get the samples of the highest purity by SDS method ,of maximum concentration by the kit method .Through com-prehensine analysis of purity and concentration we reach the conclusion that the kit method （ improved） is the best method , followed by the SDS and CTAB （ improved ） .