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

Quantitative detection method for purity of fragrant rice based on molecular biology and detection kit thereof

A quantitative detection and kit technology, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of poor accuracy and difficulty in quantitative detection of fragrant rice content, and avoid human judgment. , The results are intuitive and objective, and the operation is convenient and fast.

Pending Publication Date: 2022-04-15
WILMAR SHANGHAI BIOTECH RES & DEV CENT
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, a variety of methods have been established to detect the aroma of rice materials, among which the chewing method and the KOH method are the most commonly used methods in the traditional breeding process, but these two methods mainly rely on the human senses to determine the strength of the aroma, accuracy Poor, more difficult to achieve quantitative detection of fragrant rice content, because the fragrance of fragrant rice is not only affected by different varieties, but also affected by external environmental factors such as climate in different years
In recent years, with the rapid development of rice functional genome and sequencing technology, the research on rice aroma genes has made great progress. The genetic basis of aroma in fragrant rice is relatively complicated, but most researchers believe that aroma is controlled by a single recessive gene of

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
  • Quantitative detection method for purity of fragrant rice based on molecular biology and detection kit thereof
  • Quantitative detection method for purity of fragrant rice based on molecular biology and detection kit thereof
  • Quantitative detection method for purity of fragrant rice based on molecular biology and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Materials and methods

[0066] 1. Materials

[0067] The seeds and rice samples of different fragrant and non-fragrant rice varieties were purchased from the market.

[0068] 2. Enzymes and Reagents

[0069] The enzymes were purchased from Kangwei Century Company and Bio-Rad Company, the reagents were purchased from Sinopharm Chemical Reagent Co., Ltd., and the fluorescent quantitative PCR instrument Bio-Rad CFX96; the primers and probes used in the experiment were synthesized by Shanghai Sangon Bioengineering Company .

[0070] 3. Experimental method

[0071] 3.1 DNA extraction from rice seeds (rice samples)

[0072] Take 20 grams of rice seeds (rice samples), grind them with a grinder, weigh 50 mg of powder into a 2 mL sample lysis tube, add 500 μL of Buffer 1, vortex for 30 seconds, incubate at 52°C for 30 minutes, and rotate at 1200 rpm; Add 500 μL of Buffer 2, vortex and mix for 30 seconds, centrifuge the mixture for 5 minutes (12000 rpm), absorb 50...

Embodiment 2

[0091] Example 2: Real-time fluorescent quantitative PCR detection of non-fragrant rice and fragrant rice

[0092] Using the extracted DNA as a template, primers badh2-7-LF1 and badh2-7-LR1, non-fragrant rice-specific probe badh2-7-22MGB-P, fragrant rice-specific probe badh2-7-VIC-P4, and The endogenous reference probe badh2-7-CY5-P5 was detected by real-time fluorescent PCR. The PCR reaction system is 20 μL, including 10 μL of 2×GoldStar Best MasterMix, 1.5 μL of badh2-7-LF1 primer (concentration of 10 μM), and 1.5 μL of badh2-7-LR1 primer (concentration of 10 μM), badh2-7-22MGB-P Probe, badh2-7-VIC-P4 probe and badh2-7-CY5-P5 probe (concentration: 10 μM) were 0.25 μL each, template DNA was 2 μL, and sterile water was added to 20 μL. In the blank control, sterile water was used instead of template DNA. Each reaction was repeated three times, and the PCR amplification program used a two-step method: pre-denaturation at 95°C for 10 min; denaturation at 95°C for 15 s; annealin...

Embodiment 3

[0095] Example 3: Analysis of quantitative detection data of different internal reference genes

[0096]Using the extracted DNA as a template, primers badh2-7-LF1 and badh2-7-LR1, non-fragrant rice-specific probe badh2-7-22MGB-P, fragrant rice-specific probe badh2-7-VIC-P4, and internal The source reference probe badh2-7-CY5-P5 was detected by real-time fluorescent PCR. The PCR reaction system is 20 μL, including 10 μL of 2×GoldStar Best MasterMix, 1.5 μL of badh2-7-LF1 primer (concentration of 10 μM), and 1.5 μL of badh2-7-LR1 primer (concentration of 10 μM), badh2-7-22MGB-P Probe, badh2-7-VIC-P4 probe and badh2-7-CY5-P5 probe (concentration: 10 μM) were 0.25 μL each, template DNA was 2 μL, and sterile water was added to 20 μL. In the blank control, sterile water was used instead of template DNA. Each reaction was repeated three times, and the PCR amplification program used a two-step method: pre-denaturation at 95°C for 10 min; denaturation at 95°C for 15 s; annealing and ...

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 provides a method for quantitatively detecting the purity of fragrant rice and a kit for quantitatively detecting the purity of fragrant rice. The method for quantitatively detecting the purity of the fragrant rice comprises the following steps: (1) extracting DNA (Deoxyribose Nucleic Acid) of a rice sample; (2) carrying out PCR (Polymerase Chain Reaction) amplification by using primers capable of amplifying differential fragment sequences of the fragrant rice and the non-fragrant rice; and 3) carrying out quantitative analysis to determine the purity of the fragrant rice in the to-be-detected rice sample. The method comprises the following steps: designing efficient and sensitive fragrant rice specific primers and probes by utilizing a specific sequence, different from non-fragrant rice, of fragrant rice, carrying out real-time fluorescent PCR amplification on sample DNA, and carrying out quantitative analysis on amplification data by setting an endogenous reference gene and a reference sample, so that the purity of the fragrant rice is quantitatively judged. According to quantitative detection of the purity of the fragrant rice based on molecular biology, the problem of quantitative identification of non-fragrant rice doped in the fragrant rice is effectively solved.

Description

technical field [0001] The invention relates to biological detection technology, in particular to a method for quantitatively detecting the purity of fragrant rice by means of molecular biology. Background technique [0002] Rice (Oryza sativa) is the staple food of more than 3 billion people in the world and is one of the most important food crops. Fragrant rice, one of the types of cultivated rice, is favored by consumers at home and abroad because of its unique fragrance, and its demand continues to increase. Fragrant rice is soft, delicious and nutritious, and its market price is 1-2 times higher than that of ordinary rice. Some lawbreakers resort to deception and mix non-fragrant rice with similar grain shape and appearance into fragrant rice to make huge profits. Therefore, how to identify the purity of fragrant rice in fragrant rice simply, accurately and quickly has always been a difficult problem that everyone pays attention to. [0003] At present, a variety of m...

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
IPC IPC(8): C12Q1/6851C12Q1/6895C12N15/11
Inventor 肖玲封莉杨华牛其文
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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