Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

PCR primers for amplification of Streptomyces, and method and kit for detecting Streptomyces

A technology of streptomyces and kits, applied in the field of microorganisms, can solve problems such as inability to quickly identify, and achieve the effects of short test period, wide application range and simple operation

Active Publication Date: 2014-03-26
ADVANCED ENERGY & ENVIRONMENTAL TECH INC
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is still necessary to obtain a pure culture of the strain, and it is not possible to quickly identify whether it contains Streptomyces from a mixed sample

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • PCR primers for amplification of Streptomyces, and method and kit for detecting Streptomyces
  • PCR primers for amplification of Streptomyces, and method and kit for detecting Streptomyces
  • PCR primers for amplification of Streptomyces, and method and kit for detecting Streptomyces

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: take pure culture genomic DNA as template and carry out PCR amplification with primers of the present invention

[0038] Step 1: Select a mixed bacterial culture containing Streptomyces and extract its genomic DNA. The extraction method refers to the bacterial genomic DNA extraction kit of TAKARA company, and the genomic DNA of the mixed bacteria is obtained.

[0039] Step 2: Using the DNA obtained in step 1 as a template, prepare a 100-200ng / μL DNA sample, and prepare the following 50μL reaction system with 10-20pmol / μL PCR primers for PCR amplification reaction:

[0040] PCR buffer: 5μL

[0041] dNTPs: 4 μL

[0042] Primer F upstream primer: 1 μL

[0043] Primer R downstream primer: 1 μL

[0044] DNA: 1 μL

[0045] Taq: 0.5 μL

[0046] wxya 2 O: 37.5 μL

[0047] Step 3: After mixing according to the above system, put the PCR tube into the PCR instrument for amplification. The PCR program is as follows:

[0048]

[0049] The PCR enzyme uses Taq...

Embodiment 2

[0051] Embodiment 2: Carry out PCR amplification with soil bacteria genome DNA as template

[0052] Step 1: Take 1 to 2 grams of soil sample and extract its total microbial DNA. The extraction method refers to DNA Recovery from Soils of Diverse Composition (JIZHONG ZHOU, 1996) to obtain the total soil DNA solution.

[0053] Step 2: Using the DNA obtained in step 1 as a template, prepare a 100-200ng / μL DNA sample, and prepare the following 50μL reaction system with 10-20pmol / μL PCR primers for PCR amplification reaction:

[0054] PCR buffer: 5μL

[0055] dNTPs: 4 μL

[0056] Primer F upstream primer: 1 μL

[0057] Primer R downstream primer: 1 μL

[0058] DNA: 1 μL

[0059] Taq: 0.5 μL

[0060] ddH2O: 37.5 μL

[0061] Step 3: After mixing according to the above system, put the PCR tube into the PCR instrument for amplification. The PCR program is as follows:

[0062]

[0063] The PCR enzyme uses Taq enzyme (TAKARA)

[0064] Step 4: Electrophoresis observation. Tak...

Embodiment 3

[0066] Embodiment 3: the stability test of primer amplification Streptomyces DNA described in the present invention

[0067] Referring to Example 1 and Example 2, a pure culture of mixed strains containing Streptomyces was selected to extract its genomic DNA.

[0068] Step 2: Using the DNA obtained in step 1 as a template, prepare a 100-200ng / μL DNA sample, and prepare the following 50μL reaction system with 10-20pmol / μL PCR primers for PCR amplification reaction:

[0069] PCR buffer: 5μL

[0070] dNTPs: 4 μL

[0071] Primer F upstream primer: 1 μL

[0072] Primer R downstream primer: 1 μL

[0073] DNA: 1 μL

[0074] Taq: 0.5 μL

[0075] wxya 2 O: 37.5 μL

[0076] Step 3: After mixing according to the above system, put the PCR tube into the PCR instrument for amplification. The PCR program is as follows:

[0077]

[0078]

[0079] The PCR enzyme uses Taq enzyme (TAKARA)

[0080] Step 4: Electrophoresis observation. Take out 5 μL of the final PCR product and mi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of microbes and discloses a pair of PCR primers for amplification of streptomyces DNA, wherein the sequence of a forward primer is shown in SEQ ID No.1, and the sequence of a reverse primer is shown in SEQ ID No.2. The invention further provides a method and a kit for detecting streptomyces. The primers provided by the invention are suitable for amplification of purified streptomyces DNA as well as for amplification of purified streptomyces DNA from mixed flora, are safe, accurate, quick and stable in amplification of streptomyces, and have high specificity and sensitivity.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a PCR primer for amplifying Streptomyces, a method and a kit for detecting Streptomyces. Background technique [0002] Streptomyces belongs to the family of Actinomycetes, and Streptomyces are widely found in soil. Since Streptomyces has the largest number of species, it is difficult to classify and identify. It is generally believed that the shape of the sporocyx, the surface structure of the spores, the color of the spores, and whether melanoids are produced in the organic medium are the most important classification indicators. Streptomyces mostly grow luxuriantly on artificial medium, and a few are plant pathogenic bacteria. However, in liquid medium, the antibiotics produced by it can inhibit the growth of Streptomyces, so liquid medium is generally not used for culture and isolation. [0003] Streptomyces has important application value, many of which are antibiotic (such as ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/04C12Q1/686C12Q2531/113
Inventor 郝纯邓诗财梅海李雪
Owner ADVANCED ENERGY & ENVIRONMENTAL TECH INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com