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Primer for detecting peach-derived component in sample, method and kit

A detection method and kit technology, applied in the field of law, to achieve the effects of strong specificity, avoiding false positives, and reliable results

Inactive Publication Date: 2012-02-08
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there are no reports at home and abroad on methods and kits that can quickly, simply, specifically and sensitively detect peach-derived components in samples such as fruit juices, food, medicines, and nutritional products

Method used

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  • Primer for detecting peach-derived component in sample, method and kit
  • Primer for detecting peach-derived component in sample, method and kit
  • Primer for detecting peach-derived component in sample, method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] This embodiment tests the extraction quality of the total DNA of a sample by using plant universal primers.

[0031] By detecting the endogenous reference gene chloroplast trn gene possessed by the plant itself, the extraction quality of the total DNA of the sample can be tested.

[0032] The general primer sequences of plant chloroplast trn gene used in this example for detection of plant-derived components in samples are SEQ ID NO: 8 and SEQ ID NO: 9.

[0033] In this example, Snowflake Pear, Huiyuan Pear Juice (Beijing Huiyuan Food and Beverage Co., Ltd., China), Haitai Pear Juice (Haitai, Korea), and Huiyuan Pear Pulp Beverage (Beijing Huiyuan Food and Beverage Co., Ltd., China) were tested. , Rumeng Pear Juice (Dahu (Tianjin) Fresh Food Juice Co., Ltd., China), Lotte Pear Juice (Lotte, Korea), Guoguang Apple, Bright Apple Juice (Bright Dairy Co., Ltd. Dairy Factory, China), Huiyuan Apple Juice (Beijing Huiyuan Food & Beverage Co., Ltd., China), Dahu Apple Juice (D...

Embodiment 2

[0036] The inventors of the present invention cloned and sequenced the intron sequence of the sweet-like protein gene of peach for the first time by PCR.

[0037] In this example, the intron sequence of the sweet-like protein gene of peach was obtained by PCR cloning and sequencing.

[0038] According to the characteristics of the intron of the sweet protein gene in different plant varieties, the upstream and downstream primers were designed to amplify peach by using the apple sweet protein gene sequence (Genbank No. AY792604). The peach PCR product was purified and recovered according to the instructions of Wizard Gel Extraction Kit (Promega, USA). According to the instructions of the TaKaRa pMD19-T Vector kit (TaKaRa, Japan), the purified product was connected to pMD19-T Vector, the connection system was 10 μL, and the reaction components were: pMD19-T Vector 1 μL, PCR product 2 μL, ddH 2 O 2 μL, Solution I 5 μL, set positive and negative controls at the same time. Place t...

Embodiment 3

[0044] This embodiment uses the primer sequence of the sweet-like protein gene of peach to specifically detect the sweet-like protein gene sequence of the peach-derived component in the sample by real-time fluorescent PCR. SYBR fluorescent dye method: SYBR Green is a highly sensitive DNA fluorescent dye, which can detect at least 20pg DNA in various analyses. SYBR Green dye has a very high affinity to double-stranded DNA, and the fluorescent signal will be enhanced by 800-1000 times after combining with the product dsDNA in the PCR reaction, and it has no inhibitory effect on enzymes commonly used in molecular biology. The SYBR Green PCR reaction system is: Faststart Universal SYBRGreen Master (Roche) 12.5 μL; upstream and downstream primers (10 μmol / L) each 0.5 μL; DNA template 5 μL (about 50 ng), make up the total system to 25 μL with sterile water. Bio-Rad iQTM5 multi-color real-time PCR instrument was used for PCR amplification and result analysis. The amplification condit...

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Abstract

The present invention relates to oligonucleotide primer for detecting a peach-derived component in a sample, and further relates to a real time fluorescence PCR detection method for detecting the peach-derived component in the sample, wherein the method comprises using the specific oligonucleotide primer for the peach-derived component in the sample. The invention further relates to a PCR detection kit for rapidly detecting the peach-derived component in the sample, wherein the kit comprises the specific oligonucleotide primer for detecting the peach-derived component in the sample through the real time fluorescence PCR detection. The invention further relates to an application of the specific oligonucleotide for the peach-derived component in detection of the peach-derived component in the sample. With adopting the PCR detection method and the PCR detection kit provided by the present invention, the peach-derived component in fruit juice, food, medicine, nourishments and other samples can be simply, rapidly, specifically and sensitively detected.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the present invention relates to oligonucleotide primers used for the detection of peach-derived components in samples, a real-time fluorescent PCR detection method for determining peach-derived components in samples, and a method for rapidly detecting peach-derived components in samples. A PCR detection kit for peach-derived components, and the application of specific oligonucleotide primers and probes for peach-derived components in a sample to detect peach-derived components in a sample. Background technique [0002] my country has abundant fruit resources, and the fruit juice processing industry has developed rapidly in recent years. With the improvement of people's living standards and health awareness, fruit juice beverage products are increasingly diversified, and various types, types and brands of fruit juice beverages emerge in an endless stream. However, driven by economic in...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64C12N15/11
Inventor 黄文胜陈颖韩建勋邓婷婷吴亚君
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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