PCR determination method for Chinese medicine or Chinese medicinal crops derived from eukaryote
An identification method and eukaryotic technology are applied in the field of PCR identification of traditional Chinese medicines and traditional Chinese medicinal materials derived from eukaryotes, and can solve the problems of short fragments, difficult amplification, DNA damage, etc., and achieve low cost, simple operation, The effect of effective identification
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Embodiment 1
[0052] Example 1 Detection of Donkey-derived Components Contained in Colloidal Chinese Medicines
[0053] 1 Materials and Reagents
[0054] Materials: Donkey-hide gelatin (produced by Shandong Dong'e Co., Ltd.) DNA extraction solution (diluted to 10 ng / μL and 1 ng / μL for use); yellow gelatin (Shandong Dong'e Co., Ltd. Institute uses cowhide boiled) DNA extraction solution (diluted to 10ng / μL and 1ng / μL for use); pig skin glue (made from pigskin in Shandong Dong’a Co., Ltd. Research Institute) DNA extract (diluted to 10ng / μL and 1ng / μL for use); donkey used as a positive control , horse, cow, pig genomic DNA extract (1ng / μL).
[0055] Reagent: PCR Premix Enzyme Premix (Ex Taq TM Version): (purchased from TAKARA, Cat.No.DRR033A); DNA fluorescent dye 10000×GelRed TM : DNA fluorescent dye (purchased from Biotium, Cat. No. 41000).
[0056] 2 detection method
[0057] Take various gel DNA extracts (10ng / μL) and various meat genome DNA extracts for use, use primers designed a...
Embodiment 2
[0064] Embodiment 2 detects the bovine-derived components contained in the gelatinous traditional Chinese medicine
[0065] 1 Materials and Reagents
[0066]Materials: Donkey-hide gelatin (produced by Shandong Dong'e Co., Ltd.) DNA extraction solution (diluted to 10 ng / μL and 1 ng / μL for use); yellow gelatin (Shandong Dong'e Co., Ltd. Institute uses cowhide boiled) DNA extraction solution (diluted to 10ng / μL and 1ng / μL for use); pig skin glue (made from pigskin in Shandong Dong’a Co., Ltd. Research Institute) DNA extract (diluted to 10ng / μL and 1ng / μL for use); donkey used as a positive control , horse, cow, pig genomic DNA extract (1ng / μL).
[0067] Reagent: PCR Premix Enzyme Premix (Ex Taq TM Version): (purchased from TAKARA, Cat.No.DRR033A); DNA fluorescent dye 10000×GelRed TM : DNA fluorescent dye (purchased from Biotium, Cat. No. 41000).
[0068] 2 detection method
[0069] Take various gel DNA extracts (1ng / μL) and various meat genome DNA extracts for use, use pr...
Embodiment 3
[0076] Embodiment 3 detects the porcine origin component contained in the gelatinous traditional Chinese medicine
[0077] 1 Materials and Reagents
[0078] Materials: Donkey-hide gelatin (produced by Shandong Dong'e Co., Ltd.) DNA extraction solution (diluted to 10 ng / μL and 1 ng / μL for use); yellow gelatin (Shandong Dong'e Co., Ltd. Institute uses cowhide boiled) DNA extraction solution (diluted to 10ng / μL and 1ng / μL for use); pig skin glue (made from pigskin in Shandong Dong’a Co., Ltd. Research Institute) DNA extract (diluted to 10ng / μL and 1ng / μL for use); donkey used as a positive control , horse, cow, pig genomic DNA extract (1ng / μL).
[0079] Reagent: PCR Premix Enzyme Premix (Ex Taq TM Version): (purchased from TAKARA, Cat.No.DRR033A); DNA fluorescent dye 10000×GelRed TM : DNA fluorescent dye (purchased from Biotium, Cat. No. 41000).
[0080] 2 detection method
[0081] Take various gel DNA extracts (1ng / μL) and various meat genome DNA extracts for use, use pr...
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