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

Method for identifying porphyra yezoensis high-temperature-resistant strain TM-18, molecular marker and construction method for same

A technology of Porphyra striae and molecular marker, which can be applied to biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., and can solve problems such as difficulty in distinguishing heat-resistant strains from other strains, limiting the promotion and application of improved varieties, etc. , to achieve the effect of simple and intuitive judgment criteria, stable test results and high polymorphism

Inactive Publication Date: 2014-11-26
SHANGHAI OCEAN UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, affected by factors such as the sea environment, it is difficult to identify high-temperature-resistant strains from other strains by traditional morphological methods, which limits the promotion and application of improved varieties.

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
  • Method for identifying porphyra yezoensis high-temperature-resistant strain TM-18, molecular marker and construction method for same
  • Method for identifying porphyra yezoensis high-temperature-resistant strain TM-18, molecular marker and construction method for same
  • Method for identifying porphyra yezoensis high-temperature-resistant strain TM-18, molecular marker and construction method for same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The method for constructing a molecular marker for identifying the high temperature resistant strain TM-18 of Porphyra variegata comprises the following steps:

[0044] (1) Extraction of total DNA: Collect fresh thallus of the high temperature-resistant strain TM-18 of P.

[0045] (2) SRAP marker analysis: use the SRAP primer set to analyze the DNA obtained in step (1). For primer sequences, see: "New molecular markers—SRAP and TRAP and their applications" (Liu Liwang, Gong Yiqin, Huang Hao, Zhu Xianwen, "Heritage" 26 (5): 777-781, 2004)

[0046]The SRAP-PCR reaction system is: 10×PCR buffer2.5μL, 20ng template DNA, forward and reverse primers 20ng each, the reaction system contains 2.0mmol / L MgCl2 (final concentration), 0.2mmol / L dNTP (final concentration) and 1U Taq DNA polymerase, plus sterile ddH 2 0 to 25 μL;

[0047] The reaction program was: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 1 min, annealing at 35°C for 1 min, extension at 72°C for 1...

Embodiment 2

[0072] The heat-resistant strain TM-18 and the non-heat-resistant strain of Porphyra zebra were identified respectively (samples 1-3 were the high-temperature-resistant strain TM-18 of P.

[0073] (1) Extraction of total DNA: the same method as in step (1) of Example 1;

[0074] (2) SCAR marker verification: use the primers designed in step (5) of Example 1 to perform PCR amplification on the obtained total DNA;

[0075] The reaction system is: 10×PCR buffer1.5μL, 20ng template DNA, forward and reverse primers 20ng each, the reaction system contains 2.0mmol / L MgCl 2 (final concentration), 0.2mmol / L dNTP (final concentration) and 1U Taq DNA polymerase, plus sterilized ddH 2 0 to 15 μL;

[0076] The reaction program was: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 52°C for 45 s, extension at 72°C for 45 s, repeating 35 cycles; extension at 72°C for 7 min.

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 belongs to the fields of molecular biology DNA (deoxyribonucleic acid) marking technology and applications, and relates to a molecular marker for porphyra yezoensis high-temperature-resistant strain TM-18 and a construction method for the same. Specifically, a porphyra yezoensis strain obtained by artificial selection and breeding is analysed by virtue of an SRAP (sequence-related amplified polymorphism) marking technology, and the specific bands of the high-temperature-resistant strain TM-18 are found via PCR (polymerase chain reaction) electrophoresis detection; and specific primers are designed via recovering and sequencing, and converted to SCAR (sequence characterized amplified regions) markers for extension and strain verification. A method for identifying porphyra yezoensis high-temperature-resistant strain TM-18 by the molecular marker comprises the following steps of: extracting the total DNA of porphyra yezoensis; performing PCR amplification by the specific primers; performing electrophoresis detection on amplification products; and performing result judgement, specifically, if amplification products with a length of 341 bp are generated, then judging that the strain is a high-temperature-resistant strain TM-18. In the method, identification can be performed only by one-time simple PCR and agarose electrophoresis; and the method is simple and rapid to operate, and free from the influence of environmental factors, and is an ideal means of strain identification.

Description

technical field [0001] The invention belongs to the field of DNA marker technology and application of molecular biology, and relates to a molecular marker of a high-temperature-resistant strain of Porphyra zebra TM-18 and a construction method thereof. Background technique [0002] Porphyra zebra is the main cultivated variety of laver in the north of the Yangtze River in my country. It occupies an important position in the economic seaweed industry, with a total output value of about 3 billion yuan. [0003] Porphyra zebra is a cold-temperate seaweed. The Porphyra filaments in the seedling stage are resistant to high temperatures and can naturally survive the high temperature period in summer and autumn. The suitable growth temperature is 20-25°C; the filaments mature in early autumn and release ascospores. Conchospores germinate and grow into fronds, which grow at low temperatures in late autumn, winter, and early spring. The optimum temperature for the growth of young alg...

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 Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 严兴洪贾威黄林彬
Owner SHANGHAI OCEAN UNIV
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