藏茴香总DNA提取方法探究
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摘要
本试验取藏茴香叶片制成干样、鲜样,采用SDS法、CTAB法、改良SDS法、改良CTAB法、试剂盒法五种提取方法对其总DNA进行提取,比较DNA样品的纯度、得率、完整度等指标,以期获得优质高效的藏茴香总DNA的提取方法。试验结果如下:材料选用藏茴香叶片干样所提取的总DNA效果好。用试剂盒法和改良CTAB法提取的藏茴香总DNA样品质量优良、完整性好、纯度高,明显优于其他方法;使用改良CTAB法和试剂盒法所提取的藏茴香DNA进行PCR检测,获得了明亮清晰的扩增图谱;从成本方面考察,试剂盒法远高于其他方法。因此,改良CTAB法提取藏茴香干样总DNA效果最佳。
In order to acquire the efficient method to extract genomic DNA from Carum carvi, the fresh and dried leaves of Carum carvi were used as experimental material in this research. The DNA was extracted by 5 different kinds of DNA extraction methods, including method of SDS, method of CTAB, method of modified SDS, method of modified CTAB, and TIANGEN DNA Extraction Kit method, and the purity, yield and integrity of DNA samples extracted by these methods were compared. The results showed that using dried leaves of Carum carvi could get higher-quality genomic DNA. In the respect of purity, yield and concentration, methods of modified CTAB and TIANGEN DNA Extraction Kit were better than others. The DNA extracted by improved CTAB method and TIANGEN DNA Extraction Kit method was used for PCR detection, and a bright and clear amplification map was obtained. In terms of cost, the TIANGEN DNA Extraction Kit method was much higher than other methods. In conclusion, the method of modified CTAB was the most suitable method for genomic DNA extraction from the dried leaves of Carum carvi.
In order to acquire the efficient method to extract genomic DNA from Carum carvi, the fresh and dried leaves of Carum carvi were used as experimental material in this research. The DNA was extracted by 5 different kinds of DNA extraction methods, including method of SDS, method of CTAB, method of modified SDS, method of modified CTAB, and TIANGEN DNA Extraction Kit method, and the purity, yield and integrity of DNA samples extracted by these methods were compared. The results showed that using dried leaves of Carum carvi could get higher-quality genomic DNA. In the respect of purity, yield and concentration, methods of modified CTAB and TIANGEN DNA Extraction Kit were better than others. The DNA extracted by improved CTAB method and TIANGEN DNA Extraction Kit method was used for PCR detection, and a bright and clear amplification map was obtained. In terms of cost, the TIANGEN DNA Extraction Kit method was much higher than other methods. In conclusion, the method of modified CTAB was the most suitable method for genomic DNA extraction from the dried leaves of Carum carvi.
引文
Hou Y.X.,Tang H.R.,Zhang Y.,Luo Y.,and Dong X.L.,2009,Comparison of the DNA extraction methods on the quality of DNA extracted from different parts of Salvia splendens,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),28(1):94-100(侯艳霞,汤浩茹,张勇,罗娅,董晓莉,2009,DNA提取方法对一串红不同部位DNA提取的比较,基因组学与应用生物学,28(1):94-100)
Lemhadri A.,Hajji L.,Michel J.B.,and Eddouks M.,2006,Cholesterol and triglycerides lowering activities of caraway fruits in normal and streptozotocin diabetic rats,Journal of Ethnopharm,106(3):321-326
Hu F.R.,Guo R.R.,Wang F.,and Wang J.Y.,2013,Choosing suitable method to extract genomic DNA from the leaves of in-tube seeding of the hybrid offspring of oriental lily,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),11(2):262-266(胡凤荣,国荣荣,王斐,王江勇,2013,东方百合杂种后代试管苗叶片总DNA提取方法的选择,分子植物育种,11(2):262-266)
Qiu J.B.,Ding W.Y.,and Chen S.B.,2012,Comparison of the efficiency of different DNA extraction methods on mangrove,Zhejiang Haiyang Xueyuan Xuebao(Journal of Zhejiang O-cean University),31(5):402-408(仇建标,丁文勇,陈少波,2012,几种DNA提取方法对红树植物秋茄叶片DNA提取效果的比较,浙江海洋学院学报,31(5):402-408)
Xiao T.T.,Zhu Y.,Ye B.P.,Jin G.Q.,and Qin M.J.,2010,Comparison of methods of DNA extraction from Iris L.,Zhong guo Yesheng Zhiwu Ziyuan(Chinese Wild Plant Resources),29(3):46-50(肖婷婷,朱艳,叶波平,金国虔,秦民坚,2010,鸢尾属药用植物总DNA提取方法的比较研究,中国野生植物资源,29(3):46-50)
Yi Q.P.,Luo Z.R.,and Zhang Q.L.,2007,Advances of genome DNA extraction and purification in plants,Anhui Nongye Kexue(Journal of Anhui Agricultural Sciences),35(25):7789-7791(易庆平,罗正荣,张青林,2007,植物总基因组DNA提取纯化方法综述,安微农业科学,35(25):7789-7791)
Lemhadri A.,Hajji L.,Michel J.B.,and Eddouks M.,2006,Cholesterol and triglycerides lowering activities of caraway fruits in normal and streptozotocin diabetic rats,Journal of Ethnopharm,106(3):321-326
Hu F.R.,Guo R.R.,Wang F.,and Wang J.Y.,2013,Choosing suitable method to extract genomic DNA from the leaves of in-tube seeding of the hybrid offspring of oriental lily,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),11(2):262-266(胡凤荣,国荣荣,王斐,王江勇,2013,东方百合杂种后代试管苗叶片总DNA提取方法的选择,分子植物育种,11(2):262-266)
Qiu J.B.,Ding W.Y.,and Chen S.B.,2012,Comparison of the efficiency of different DNA extraction methods on mangrove,Zhejiang Haiyang Xueyuan Xuebao(Journal of Zhejiang O-cean University),31(5):402-408(仇建标,丁文勇,陈少波,2012,几种DNA提取方法对红树植物秋茄叶片DNA提取效果的比较,浙江海洋学院学报,31(5):402-408)
Xiao T.T.,Zhu Y.,Ye B.P.,Jin G.Q.,and Qin M.J.,2010,Comparison of methods of DNA extraction from Iris L.,Zhong guo Yesheng Zhiwu Ziyuan(Chinese Wild Plant Resources),29(3):46-50(肖婷婷,朱艳,叶波平,金国虔,秦民坚,2010,鸢尾属药用植物总DNA提取方法的比较研究,中国野生植物资源,29(3):46-50)
Yi Q.P.,Luo Z.R.,and Zhang Q.L.,2007,Advances of genome DNA extraction and purification in plants,Anhui Nongye Kexue(Journal of Anhui Agricultural Sciences),35(25):7789-7791(易庆平,罗正荣,张青林,2007,植物总基因组DNA提取纯化方法综述,安微农业科学,35(25):7789-7791)