骨胶原蛋白肽粉中DNA的提取及猪、羊源性成分的检测

Extraction of DNA from bone collagen peptide powder and detection of pig and sheep derived components

中文关键词:  骨胶原蛋白肽  DNA提取  实时荧光PCR  源性成分检测

英文关键词:bone collagen peptide  DNA extraction  real-time fluorescence PCR  source component detection

中文摘要:

      目的 优化深加工骨胶原蛋白肽粉中DNA的提取方法, 鉴定检测其动物源性成分。方法 采用2种试剂盒法, 并对试剂盒部分步骤进行适度修改后提取骨胶原蛋白肽粉中的基因组DNA, 分别用猪、羊引物和探针对提取的DNA片段进行实时荧光PCR扩增检测。结果 只有DNeasy Blood & Tissue Kit提取试剂盒法提取的DNA纯度较好, A260/A280比值为1.73, 浓度为8.2 ng/μL, 并且经18S rDNA片段的PCR扩增结果表明样品中含有哺乳动物DNA, 且DNA中不含抑制PCR反应物质。检测结果表明该骨胶原蛋白肽粉中含有猪源性成分。 结论 采用试剂盒法提取深加工骨胶原蛋白肽粉中DNA时, 需对部分步骤做修改后才可进行。

英文摘要:

      Objective To optimize the method for extracting DNA from deep processed bone collagen peptide powder, and to identify and detect animal-derived components. Methods Totally 2 kit methods were used to extract genomic DNA from the collagen peptide powder after some steps of the kit were moderately modified. The extracted DNA fragments were amplified by real-time fluorescence PCR using primers and probes from pigs and sheep respectively. Results Only the DNA extracted by the DNeasy Blood & Tissue Kit extraction kit method had good purity. The A260/A280 ratio was 1.73 and the concentration was 8.2 ng/μL, and the PCR amplification results of the 18S rDNA fragment showed that the sample contained mammalian DNA, and the DNA did not contain a substance that inhibited the PCR reaction. Test results showed that the collagen peptide powder contained pig-derived ingredients. Conclusion When the kit method is used to extract DNA from deep-processed collagen peptide powder, some steps need to be modified before proceeding.