DNA barcode primers, DNA barcodes, kit and method for rapidly identifying Blastobotrys adeninivorans strain, and applications of DNA barcode primers and DNA barcodes

A technology of eating Arthrophthora adenine and barcode primers, applied in the directions of DNA/RNA fragments, recombinant DNA technology, microorganism-based methods, etc., can solve the problems of lack of Pu'er tea, damage to the economic interests of Pu'er tea production enterprises, etc., to improve identification Accuracy, Ensuring Health and Prosperity, Low-Required Effects

Active Publication Date: 2018-06-05
MENGHAI TEA IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, due to the increasing market demand, many small-scale manufacturers lack systematic and in-depth research on Pu-erh tea, and wantonly abuse many other people's patents for profit. A series of methods, such as Pu’er tea processing and production, have greatly damaged the economic interests of some Pu’er tea production enterprises with relevant patents.

Method used

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  • DNA barcode primers, DNA barcodes, kit and method for rapidly identifying Blastobotrys adeninivorans strain, and applications of DNA barcode primers and DNA barcodes
  • DNA barcode primers, DNA barcodes, kit and method for rapidly identifying Blastobotrys adeninivorans strain, and applications of DNA barcode primers and DNA barcodes
  • DNA barcode primers, DNA barcodes, kit and method for rapidly identifying Blastobotrys adeninivorans strain, and applications of DNA barcode primers and DNA barcodes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1: Based on the homology comparison of the sequences amplified with the first, second, and third primer pairs to determine whether the strain to be tested is Arthrospora adenophagous TMCC 70007

[0069] 1. Source of strains: See Table 1 below for information on the selected strains:

[0070] Table 1

[0071]

[0072] In the above table 1, the TMCC 70007 strain was preserved in the China General Microbiological Culture Collection Center (CGMCC) with the preservation number CGMCCNo.8683; the CBS strains were all purchased from the CBS-KNAW Fungal Biodiversity Center, the Netherlands .

[0073] 2. Extract the strain DNA separately: OMEGA E.Z.N.A. TM Yeast DNA Kit D3370-01 (manufactured by OMEGA) was used to extract yeast genomic DNA, and the DNA concentration of the sample was diluted to 0.5 μg / μL with sterilized deionized water.

[0074] 3. To amplify the DNA fragments, use the following 3 pairs of primers respectively to carry out the polymerase chain reac...

Embodiment 2

[0089] Example 2: Using the Phylogenetic Tree Method to Establish Identification Rules to Identify Species

[0090] Using the steps described in Example 1 to obtain the test sequence of the test strain, the TMCC 70007, CBS 8244 T , CBS 8335, CBS 7350 strains and CBS 7370 and CBS 6800 of the species Blastobotrysraffinosifermentans T The strains to be tested were merged using MEGA 5.10 to construct a Neighbor-joining phylogenetic tree based on the Kimura-2-parameter model. The method was the Bootstrap method, and the Bootstrap repeat No. was 1000. If the tested sequence of the tested strain is clustered with the standard sequence of TMCC 70007 (that is, the same branch and the same evolutionary distance), it means that the tested strain is the Pu-erh tea fermentation strain TMCC 70007.

[0091] Such as figure 1 Shown, TMCC 70007, CBS 8244 of Arthrospora adenophagous species T , CBS 8335, CBS 7350 strains and CBS 7370 and CBS 6800 of the species Blastobotrys raffinosifermentan...

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PUM

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Abstract

The invention belongs to the field of species and strain identification, and particularly relates to a DNA barcode primer composition, a DNA barcode composition, a kit and a method for identifying a Blastobotrys adeninivorans strain, and applications of the DNA barcode primer composition and the DNA barcode composition, wherein the DNA barcode composition comprises a first DNA barcode representedby SEQ ID No.1, a second DNA barcode represented by SEQ ID No.2, and a third DNA barcode represented by SEQ ID No.3, and the first DNA barcode, the second DNA barcode and the third DNA barcode are derived from the genome of a Blastobotrys adeninivorans TMCC 70007 strain. According to the present invention, the DNA barcodes can quickly identify the Puer fermentation strain Blastobotrys adeninivorans TMCC 70007 and can rapidly and accurately identify the Blastobotrys adeninivorans TMCC 70007 from easily confused strain or other strains in the same species.

Description

technical field [0001] The invention belongs to the field of identification of species and strains, and in particular relates to a DNA barcode primer composition, a DNA barcode composition, a kit, a method and an application for identifying Arthrospora adenorivorum strains. 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. Pu'er tea has also attracted people's attention because of its health effects such as weight loss, lowering blood sugar and blood lipids, preventing and improving cardiovascular diseases, anti-aging, anti-cancer, anti-inflammatory, aiding digestion and nourishing the stomach. In recent years, Pu'er tea has become more and more popular, and the increasing market demand has driven the development of Pu'er tea industry and promoted the economic growth of Yunnan. [0003] Pu'er tea is a large-leaf ferment...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895
Inventor 田飞唐蜀昆职晓阳高林瑞徐平施佳辉高慧英丁章贵
Owner MENGHAI TEA IND
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