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Primer probe and method for detecting cocoa-derived components in cocoa powder

A cocoa-derived, primer-probe technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. Reliable results, simple operation, avoid false positive effect

Pending Publication Date: 2022-04-15
PONY TESTING INT GRP SHANGHAI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no method for rapid, specific and sensitive detection of cocoa-derived components in cocoa powder at home and abroad

Method used

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  • Primer probe and method for detecting cocoa-derived components in cocoa powder
  • Primer probe and method for detecting cocoa-derived components in cocoa powder
  • Primer probe and method for detecting cocoa-derived components in cocoa powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] In this example, the specificity and sensitivity of cocoa primer pairs and probes are evaluated through the following tests.

[0022] The specificity and detection sensitivity of the cocoa primer-probe combination can be determined by detecting the microsatellite DNA sequence. The reaction system is: Fast Start Universal PCR Master Mix 12.5 μL; probe (10 μM) 0.5 μL; upstream and downstream primers (10 μM) 0.5 μL; template DNA 5 μL; add sterile water to a total volume of 25 μL. The reaction program was 95°C for 10 min; 95°C for 15 s; 60°C for 1 min, 40 cycles.

[0023] The primers and probe sequences used to detect cocoa are:

[0024] The primer sequences are SEQ ID No.1 and SEQ ID No.2, the probe sequence is SEQ ID No.3, a fluorescent quenching group TAMRA is connected to the 3' end, and a fluorescent reporter group FAM is connected to the 5' end.

[0025] The main detection instruments used:

[0026] Micropipette (10μL, 100μL, 1000μL), fluorescent quantitative PCR i...

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Abstract

The invention relates to a real-time fluorescent PCR (polymerase chain reaction) detection method for qualitatively detecting cocoa-derived components in cocoa powder. The method comprises the step of using specific oligonucleotide primers and fluorescent labeled probes aiming at the cocoa components in the cocoa powder. The invention also relates to application of the specific oligonucleotide primer and the fluorescent probe aiming at the cocoa component in the cocoa powder in qualitative detection of the cocoa-derived component. By using the real-time fluorescent PCR detection method disclosed by the invention, the cocoa-derived components in the cocoa powder can be accurately and sensitively determined.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the present invention relates to oligonucleotide primers and fluorescently labeled probes for the detection of cocoa-derived components, as well as a real-time fluorescent PCR detection method and a sample for determining cocoa-derived components Use of oligonucleotide primers and probes specific for cocoa-derived components in the detection of cocoa-derived components. Background technique [0002] Cocoa powder is a cocoa product processed directly from cocoa beans. Cocoa powder has a unique fragrance, strong fragrance, rich in carbohydrates, cocoa butter, polyphenols, protein and other nutritional elements and active ingredients, and is an important raw material for making chocolate. With the rapid development of the cocoa processing industry, the adulteration of counterfeit or inferior cocoa powder exists in domestic and foreign markets, which not only affects the health and intere...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
Inventor 韩建勋宋敬宁侯卫静
Owner PONY TESTING INT GRP SHANGHAI CO LTD
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