大体积肉中DNA提取方法的优化

Optimization of DNA extraction method from large volume of meat

中文关键词:  DNA提取  大体积肉类组织  试剂盒

英文关键词:DNA extraction  large volume of meat  kit

中文摘要:

      目的 优化一种从大体积肉类中提取DNA的方法。方法 从大组织肉上多点取样, 用组织粉碎机打碎, 称取10 g肉加入90 mL PBS缓冲液和450 μL蛋白酶K, 60 ℃消化至澄清后, 混匀, 取200 μL消化液用试剂盒进行DNA提取; 再与常规试剂盒方法提取的DNA在同条件进行实时荧光PCR及琼脂糖凝胶电泳验证。结果 2种方法提取DNA的Ct值差异在0.5个循环以内, 无显著性差异(P>0.05); 电泳结果显示, 2种方法所扩增的为同一产物。结论 该方法取样具有代表性, 可适用于大体积肉类掺假检测或动物源性成分检测中的DNA提取。

英文摘要:

      Objective To optimize a method of extracting DNA from large volume of meat. Methods Using multipoint sampling from large organization of meat and tissue destrustor for crushing, 10 g meat was weighed, added 90 mL PBS buffer and 450 μL prolease K, mixed evenly after digesting until clarification at 60 ℃, and then 200 μL digestive solution was taken for DNA extraction with kit. Meanwhile, common kit method was also used to extract DNA. Then DNA extracted with these two different methods were verified by real-time PCR and agarose gel electrophoresis. Results The difference of Ct values of DNA extracted with two different methods was not significant (P>0.05) with the variances within 0.5 cycle. The electrophoresis results showed that two methods obtained the same amplification products. Conclusion This established method ensures that the sampling was more representative, which can be applied for meat adulteration detection or DNA extraction in the detection of animal derived ingredients.