DNA barcode, primers, kit, method, and applications of DNA barcode, primers and the kit
A barcode and primer pair technology, applied in the field of DNA barcodes, primers, and kits, can solve problems such as damage to the economic interests of Pu'er tea production enterprises, lack of Pu'er tea, etc., and achieve rapid and accurate identification and distinction, low completeness requirements, and specificity. Good results
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Embodiment 1
[0074] Embodiment 1: Obtaining of SOF1 gene and DNA barcode
[0075] 1. Using the high-coverage proteome technology, pFind and pAnno software were used to conduct a deep coverage study of the proteome of the industrial fermentation fungus Arthrospora adenorivorum TMCC 70007, and its genome was verified for its annotated coding genes. Specifically, in order to discover new protein coding regions, using the Six Frame Translation strategy in systematic proteomics, a six-frame translation database of the genome data of Arthrospora adenophagous TMCC 70007 was obtained, exhaustively enumerating There are 6 coding possibilities of the genome (+1, +2, +3, -1, -2, -3), and the protein sequence is called "six-box translation protein sequence", and the corresponding nucleic acid sequence is called "six-box translation Nucleic acid sequence". Generally speaking, a six-frame translated nucleic acid sequence is the sequence from one terminator to the next, which is also referred to herein ...
Embodiment 2
[0099] Example 2. Identification of bacterial strains using DNA barcodes
[0100] According to the amplification result of the sample to be tested and the sequence homology with Arthrospora adenophagous TMCC 70007 strain SEQ ID NO.1, it is determined whether the sample to be tested is the industrially applied strain of Arthrospora adenorivorum TMCC 70007 .
[0101] (1) Based on SEQ ID NO.1, use the NCBI primer design tool to design PCR primers at both ends of the gene. The amplified product must include the start and end sites of the gene, and the sequences of the forward and reverse primers are SOF1-F: 5 '-GCCGCACGTCCAATATTTTTC-3'; SOF1-R: 5'-GCTGATCGGGTAGAGCAAGT-3'.
[0102] Figure 8 The position of the primer used to amplify SEQ ID NO.1 in one embodiment of the present invention is shown, the underlined region in the sequence is the region where the primer is located, and the ATG and TAG in bold font are the start and stop sites, The gray background region is the intron r...
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