不同提取方法提取辣椒及其制品中DNA效果研究

摘要

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.%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.