PCR identification method of Chinese medicament turtle-derived component
A technology of source components and identification methods, applied in the field of PCR identification of tortoise-derived components in traditional Chinese medicine, can solve the problems of poor practicability, less DNA content, cumbersome operations, etc., and achieve the effects of low cost, effective identification, and simple operation
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Embodiment 1
[0037] Example 1 Detection of turtle-derived components contained in various meat genomes
[0038] 1 Materials and Reagents
[0039] Materials: Genomic DNA extracts (1ng / μL) from various meats including turtles, donkeys, horses, cattle, and pigs.
[0040] Reagent: PCR premix enzyme Premix Ex-Taq TM Hot Start Version: (purchased from TAKARA, Cat.No.DRR030A); DNA fluorescent dye 10000×GelRed TM : DNA fluorescent dye (purchased from Biotium, Cat. No. 41000).
[0041] 2 detection method
[0042] The genomic DNA extracts of various meats were taken for use, and PCR reactions were performed using primers designed according to the SINE sequence Pol III / SINE of Cryptocervica, and the primers were dissolved in sterile water to a concentration of 10 μM for use. The specific loading conditions of the 25 μL reaction system are as follows:
[0043] Premix Ex-Taq TM Hot Start Version 12.5 μL, Primer Tortoise up 0.5 μL, Primer Tortoisedown 0.5 μL, Genomic DNA Extract 1 μL, Sterile do...
Embodiment 2
[0048] Embodiment 2 detects the tortoise-derived component contained in the glue class traditional Chinese medicine
[0049] 1 Materials and Reagents
[0050] Materials: tortoise shell glue (produced by Shandong Dong'e Co., Ltd.) DNA extraction solution (10ng / μL); donkey-hide gelatin (produced by Shandong Dong'e Co., Ltd.) DNA extraction solution (10ng / μL); yellow gelatin (Shandong Dong'e Co., Ltd. The research institute uses cowhide boiled) DNA extraction solution (10ng / μL); pig skin glue (Shandong Dong'a Co., Ltd. research institute uses pigskin boiled) DNA extraction solution (10ng / μL).
[0051] Reagent: PCR premix enzyme Premix Ex-Taq TM Hot Start Version; (purchased from TAKARA, Cat.No.DRR030A); DNA fluorescent dye 10000×GelRed TM : DNA fluorescent dye (purchased from Biotium, Cat. No. 41000).
[0052] 2 detection method
[0053] Take various gel DNA extracts (10ng / μL) for use, and use primers designed according to the SINE sequence PolIII / SINE of Cryptocervica for P...
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