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研究了一种可有效排除细胞内多酚类等杂质污染的简便、快捷、经济的果树基因组DNA提取方法.其改良策略是:在细胞核被裂解前去除细胞质中的多酚类等杂质,防止多酚类物质被氧化,从而排除多酚类等杂质的干扰.利用这一方法提取的柑桔、枇杷、梨、苹果、湖北海棠、桃、樱桃等7种果树基因组DNA均能被限制性内切酶完全消化,获得理想的Southern杂交结果,也可作为PCR模板应用,成功地进行特异性基因扩增和RAPD.
A simple, rapid and economical method for the extraction of genomic DNA from fruit trees was studied, which can effectively eliminate the contamination of polyphenols and other impurities in cells. The improved strategy is: remove the cytoplasm of polyphenols and other impurities before the nucleus is cracked to prevent the polyphenols are oxidized, thus eliminating the interference of polyphenols and other impurities. Genomic DNA of seven kinds of fruit, such as citrus, loquat, pear, apple, Hubei begonia, peach and cherry, extracted by this method can be completely digested by restriction enzyme and obtained ideal Southern hybridization result, Application, successful gene specific amplification and RAPD.