Molecular internal standard substance for identifying authenticity of objects and application of molecular internal standard substance
A technology for internal standards and articles, which is applied in the field of molecular internal standards for identifying authenticity of articles, can solve problems such as troublesome design of amplification primers and false positive results, and achieve the effect of improving specificity
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[0036] Two nucleotide sequences B1 and B-1 are designed as molecular internal standards, wherein the sequences B1 and B-1 are as follows:
[0037] B1:
[0038]
[0039] B-1:
[0040]
[0041] The underlined part of the sequence B1 and B-1 is a palindromic sequence that can form a hairpin structure. For example, the structure of the sequence B1 is as follows;
[0042] TATAGC-5'
[0043]
[0044] The wavy sequence parts of the sequences B1 and B-1 cannot form a hairpin structure, which contains an endonuclease cutting site (AATT), and the wavy sequence parts of B1 and B-1 can be complementary paired.
[0045] Sequences B1 and B-1 contain two restriction endonuclease cutting sites BstUI (5'-CGCG-3') and Tsp509I (5'-AATT-3'), and the amplified products can be digested by these two Digested by endonucleases.
[0046] The endonuclease used in the embodiment of the present invention is Vent (exo-) DNA polymerase, which is a thermophilic polymerase and has strand displaceme...
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