dna barcodes, primers, kits, methods and applications
A barcode and primer pair technology, applied in the fields of DNA barcodes, primers, and kits, can solve the problems of damaging the economic interests of Pu’er tea production enterprises and the lack of Pu’er tea, and achieve rapid and accurate identification and differentiation, low integrity requirements, and easy comparison right effect
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Publication Date
- 2021-08-13
Smart Images
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Abstract
Description
technical field
[0001] The invention belongs to the field of species and bacterial strain identification, and specifically relates to DNA barcodes, primers, kits, methods and applications. Background technique
[0002] Pu-erh tea is a post-fermented tea with the geographical indication of Yunnan. It uses large-leaf sun-dried green tea as raw material and is made through a series of processes. The traditional production process of Pu'er tea is as follows: the picked fresh tea leaves are rolled, sun-dried, impurity-removed, tide-watered, heaped, dried, sieved, pressed and molded, and packaged for delivery. In the production of Pu-erh tea, the heaping fermentation process is the main factor to form the quality of Pu-erh tea. During this process, the internal components of the tea such as tea polyphenols, caffeine and some polysaccharides have undergone tremendous changes, making Pu-erh tea The special flavor, taste, quality and various health benefits of tea.
[0003] In the ...
Examples
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 Arthrospora adenophagous TMCC 70007 genome data was obtained, exhaustively enumerated 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 tran...
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 the Arthrospora adenosaurosa TMCC 70007 strain SEQ ID NO.1, it is determined whether the sample to be tested is the industrially applied strain of Arthrospora adenorivorum TMCC70007.
[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 ...