PCR determination method for Chinese medicine or Chinese medicinal crops derived from eukaryote
An identification method and eukaryotic technology, which is applied in the field of PCR identification of traditional Chinese medicine and Chinese medicinal materials derived from eukaryotic organisms, can solve the problems of short fragments, DNA damage, difficult amplification, etc., and achieve low cost, effective identification, Simple operation effect
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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 Taq (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 design...
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 Taq (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, us...
Embodiment 3
[0075] see results figure 2 , because the primers are designed according to the SINE sequence of ruminants, lane 4 has a positive band, and lane 7 is yellow gelatin, which is boiled from cowhide, so it also has a positive band. It can be seen that although the DNA content obtained from glue-like traditional Chinese medicine is very small , using the primers designed for the SINE sequence of ruminants, the bovine-derived components mixed in glue products can be detected by PCR, but no positive bands are amplified in other samples, indicating that there is no mixed cowhide in donkey-hide gelatin. Since the pair of primers is designed based on the SINE sequence of ruminants, this method can also be used to detect whether other ruminant-derived components such as sheep are mixed in glue products. Embodiment 3 detects the porcine origin component contained in the gelatinous traditional Chinese medicine
[0076] 1 Materials and Reagents
[0077] Materials: Donkey-hide gelatin (pr...
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