DNA barcode, primers, kit and method, and applications thereof
A barcode and primer pair technology, applied in the field of kits, DNA barcodes and primers, can solve the problems of lack of Pu'er tea, damage to the economic interests of Pu'er tea production enterprises, etc. Effect
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Embodiment 1
[0072] Example 1: Obtaining of LipB gene and DNA barcode
[0073] The proteome and genome analysis methods and mass spectrometry methods used in this example are all known methods in the art, so the specific operation process will not be repeated.
[0074] 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-...
Embodiment 2
[0093] Example 2. Identification of bacterial strains using DNA barcodes
[0094] 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 .
[0095] (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 respectively LipB-F: 5 '-AACCGGTCTCCGTCAAACTC-3'; LipB-R: 5'-AAGATCTCCGCGACAAGCAA-3'. For primer locations see Image 6 , the theoretical amplification length of the pair of primers is 1579bp, and the amplified sequence is SEQ ID NO.7.
[0096] (2) Source of strain
[0097] Table 4 Selected relevant strain information
[0098]
[0099] (3) Separately extract strain DNA: OMEGA E...
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