快速提取鸭血基因组DNA方法的研究

Research on Rapid Extraction of Duck Blood Genomic DNA Method

目的:以鸭血为原料,筛选出高质量、高效率的DNA提取方法。方法:选用4种不同方法提取样品DNA,并建立实时荧光PCR实验,以此鉴别掺假伪劣产品。通过研究DNA质量浓度、提取时间和扩增效果,选取高质量、高效率的提取方法。结果:苯酚/氯仿抽提法提取浓度高、成本低,但步骤烦琐、耗时长,易接触有毒有害试剂;试剂盒法提取DNA纯度较高,但价格昂贵,不适用于大批量样品;核酸提取仪操作简便,但前处理耗时长,适用于大批量产品的检验检测;细胞裂解法操作简单、快速、成本低,因未经后续纯化,所得到的DNA纯度相对较低,但同样满足PCR扩增的检测要求,适用于大批量监督抽查任务。结论:对于大批量的鸭血源性成分检测推荐使用细胞裂解法,但一旦出现阳性样品需要进行复测则需要使用不同的提取方法,以确保实验的准确性。

Objective:To screen high-quality and efficient DNA extraction methods using duck blood as raw material.Method:Four different methods were used to extract sample DNA and real-time fluorescence PCR experiments were established to identify adulterated and inferior products.Optimize high-quality and efficient extraction methods by studying DNA mass concentration,extraction time,and amplification effect.Result:The phenol/chloroform extraction method has high extraction concentration and low cost,but the steps are cumbersome,time-consuming,and prone to contact with toxic and harmful reagents;the reagent box method has high extraction quality,but it is expensive and not suitable for large batches of samples;the nucleic acid extractor is easy to operate,but the pre-treatment time is long,making it suitable for the inspection and detection of large quantities of products;the cell lysis method is simple,fast,and cost-effective.Due to the lack of subsequent purification,the purity of the obtained DNA is relatively low,but it also meets the detection requirements of PCR amplification and is suitable for large-scale supervision and sampling tasks.Conclusion:It is recommended to use cell lysis method for the detection of duck blood derived components in large quantities.However,if a positive sample occurs and retesting is required,different extraction methods need to be used to ensure the accuracy of the experiment.