不同提取方法提取辣椒及其制品中DNA效果研究
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摘要
采用CTAB法、SDS法、吸附柱法对辣椒及其制品中的基因组DNA进行提取,分别通过核酸蛋白测定仪、琼脂糖凝胶电泳、实时荧光PCR扩增检测。结果表明,CTAB法、SDS法、吸附柱法均能从鲜辣椒、干辣椒、油辣椒、辣椒酱中抽提得到DNA,从核酸蛋白测定仪检测得到的DNA提取物OD260/OD280值可看出,CTAB法和SDS法提取的DNA纯度较吸附柱法的低;DNA样品的琼脂糖凝胶电泳发现,各样品均无大范围明显拖尾,说明DNA提取完整度较好;实时荧光PCR检测分析3种提取方法的不同样品基因组DNA均出现PCR扩增曲线。经试验比较确定,提取方法的效率由高到低依次为吸附柱法>SDS法>CTAB法。可见,吸附柱法最适用于辣椒及其制品中核酸提取。
CTAB,SDS and DNA adsorption column method were adopted to extract DNA from chilli and its products.Nucleic acidprotein detector method,agarose gel electrophoresis method and real-time fluorescence PCR amplification methods were applied to measure the DNA samples.The results showed that DNA could be extracted by CTAB,SDS and adsorption column from chilli and its products.It could be seen from the OD260/OD280 value of DNA extract by nucleic acid protein detector,the concentration of DNA extracted by CTAB and SDS was lower than that by adsorption column.Agarose gel electrophoresis measurement showed DNA extracted with 3 methods there was no obvious tail smearing in all areas,the results showed that DNA had better extraction integrity.The PCR amplification curve of genomic DNA of 3 different extraction methods was detected by real-time fluor-escence PCR.Through experiment and comparison,it was determined that the efficiency of the extraction method was as following :adsorption column method >SDS method>CTAB method.In summary,the adsorption column method is the most suitable for extracting nucleic acid from chilli and its products.
CTAB,SDS and DNA adsorption column method were adopted to extract DNA from chilli and its products.Nucleic acidprotein detector method,agarose gel electrophoresis method and real-time fluorescence PCR amplification methods were applied to measure the DNA samples.The results showed that DNA could be extracted by CTAB,SDS and adsorption column from chilli and its products.It could be seen from the OD260/OD280 value of DNA extract by nucleic acid protein detector,the concentration of DNA extracted by CTAB and SDS was lower than that by adsorption column.Agarose gel electrophoresis measurement showed DNA extracted with 3 methods there was no obvious tail smearing in all areas,the results showed that DNA had better extraction integrity.The PCR amplification curve of genomic DNA of 3 different extraction methods was detected by real-time fluor-escence PCR.Through experiment and comparison,it was determined that the efficiency of the extraction method was as following :adsorption column method >SDS method>CTAB method.In summary,the adsorption column method is the most suitable for extracting nucleic acid from chilli and its products.
引文
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[5]周春阳,谢立波,李景富,等.辣椒叶片基因组DNA提取方法的研究[J].辣椒杂志,2011(3):35-37.
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[7]陈昆松,李方,徐昌杰,等.改良CTAB法用于多年生植物组织基因组DNA的大量提取[J].遗传,2004(4):529-531.
[8]桑维维,高贵珍,张兴桃,等.辣椒基因组DNA两种提取方法的比较[J].宿州学院学报,2012(8):21-24.
[9]王艳,李韶山,刘颂豪.植物总DNA样品的快速制备[J].激光生物学报,2000(1):79-81.
[10]朱英,陶刚,刘作易,等.普遍适用于农作物基因组DNA的提取方法及值得注意的几个问题[J].贵州农业科学,2005(5):20-21.
[2]田娅红,张鼎华.植物转基因研究进展及安全性分析[J].廊坊师范学院学报(自然科学版),2014(4):89-93.
[3]张虎平,牛建新,马兵钢,等.核桃DNA的提取及RAPD体系的优化[J].石河子大学学报(自然科学版),2003(4):267-270.
[4]刘萍,代红军,张立杰,等.DNA提取方法与植物种类的效应比较[J].宁夏农林科技,1998(1):15-18.
[5]周春阳,谢立波,李景富,等.辣椒叶片基因组DNA提取方法的研究[J].辣椒杂志,2011(3):35-37.
[6]孙晓东,李小丹,郭晏海,等.经济植物DNA提取与纯化的三种方法比较[J].陕西教育学院学报,2003(2):93-94.
[7]陈昆松,李方,徐昌杰,等.改良CTAB法用于多年生植物组织基因组DNA的大量提取[J].遗传,2004(4):529-531.
[8]桑维维,高贵珍,张兴桃,等.辣椒基因组DNA两种提取方法的比较[J].宿州学院学报,2012(8):21-24.
[9]王艳,李韶山,刘颂豪.植物总DNA样品的快速制备[J].激光生物学报,2000(1):79-81.
[10]朱英,陶刚,刘作易,等.普遍适用于农作物基因组DNA的提取方法及值得注意的几个问题[J].贵州农业科学,2005(5):20-21.