SpecIdent Porcine-derived component identification kit (probe method)

Product Code: AG11753

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¥1,200

Product Description

This product is a rapid identification of swine using the probe method.*1 Kit for components of animal origin. The identification of animal-derived components is based on interspecies 16S rRNA (16S ribosomal RNA) gene polymorphisms. This product contains three different fluorescent moieties (FAM, VIC, ROX) labeled probes, which allow simultaneous detection of porcine 16S rRNA gene in the same reaction tube.*2The eukaryotic 18S rRNA (18S ribosomal RNA) gene was used as a key component of the eukaryotic gene.*3 and Internal Control, which allows you to monitor whether a reaction is proceeding properly. Detection of the 18S rRNA gene allows monitoring of the quality of eukaryotic genomic DNA extraction.
The 2X Master Mix for Porcine is already mixed with DNA polymerase, dNTP Mixture and reaction buffer, so it is very simple to prepare the reaction solution, just add the Primer Mix for Porcine, Probe Mix for Porcine, RNase free water, and RNase free water to the sample to be tested. It is easy to prepare the reaction solution, just add the Primer Mix for Porcine, Probe Mix for Porcine, RNase free water and the sample to be examined.
The PCR reaction system of this product has been optimized with the use of a superior reaction performance Pro Taq The HS system (mixed with Taq antibody) can effectively inhibit non-specific amplification, and together with the carefully optimized buffer, it improves PCR amplification efficiency and enables highly sensitive Real Time PCR reactions.
* 1: This product can detect common pig breeds, such as common pig ( Sus domesticus ), certain breeds of pigs may not be accurately detected because of possible sequence variation in different breeds.
*2: The 16S rRNA gene in this product can specifically detect pig-derived components, and the primer probe sequences are referenced from the Agricultural Industry Standard of the People's Republic of China NY/T 3309-2018.
*3: The 18S rRNA gene in this product can specifically detect eukaryotic organisms, and the primer probe sequences are referenced from the National Standard of the People's Republic of China GB/T 38164-2019.

Product Advantages

① This product combines DNA polymerase, dNTP Mixture and reaction buffer into one tube, which is easy and quick to operate, and can effectively reduce the risk of sample contamination caused by experimental operations and shorten the time required for experiments.
②This product utilizes a superior performance Pro Taq HS and optimized reaction system with high specificity and amplification efficiency.
③ This product can simultaneously detect pig-derived 16S rRNA gene, eukaryotic 18S rRNA gene and Internal Control in one tube, and can specifically identify pig-derived components.

Product Composition
individual parts making up a compound norm
2X Master Mix for Porcine*1 625 μl
Primer Mix for Porcine*2 50 μl
Probe Mix for Porcine*3 50 μl
Control DNA for Porcine*4 25 μl
RNase free water 1 ml

*1: 2X Master Mix for Porcine contains Pro Taq HS DNA Polymerase, dNTP Mixture and reaction buffer, etc.
*2: Primer Mix for Porcine contains the porcine 16S rRNA gene, the eukaryotic 18S rRNA gene, primers for the Internal Control assay and the Internal Control template.
*3: Probe Mix for Porcine contains porcine-derived 16S rRNA gene, eukaryotic 18S rRNA gene, and probes for Internal Control assay, and should be stored under light.
*4: Control DNA for Porcine is a plasmid template containing only sequences specific to porcine-derived components.

Preservation and transportation
Storage temperature: -20℃ storage
Transportation temperature: dry ice transportation or -20℃ ice bag transportation

Porcine gDNA was triple-tested for the porcine 16S rRNA gene (FAM channel), the eukaryotic 18S rRNA gene (ROX channel), and the Internal Control gene (VIC channel) using the ABI QuantStudioTM 5 Real-Time PCR System. The quantification instrument used was the ABI QuantStudioTM 5 Real-Time PCR System, and the experimental groups are listed below:

Group Template
Sample 1 10 ng Porcine gDNA
Sample 2 1 ng Porcine gDNA
Sample 3 100 pg Porcine gDNA
Sample 4 10 pg Porcine gDNA
Sample 5 1 pg Porcine gDNA
Negative Control Replace template with RNase free water
Positive Control Control DNA for Porcine

1) The results of the Sample group and the Negative Control group are as follows:



The results of pig-derived 16S rRNA detection are shown above:
1. The amplification efficiency was 99%, R2= 0.9977;
2. Accurate quantification can be performed over a wide template range, and the amplification curve of qPCR reaction with 10 ng-1 pg gDNA shows good linearity;
3. Negative Control was not detected within 45 cycles.



Eukaryotic 18S rRNA assay results are shown above:
1, with an amplification efficiency of 103%, R2= 0.9982;
2. Accurate quantification can be performed over a wide template range, and the amplification curve of qPCR reaction with 10 ng-1 pg gDNA shows good linearity;
3. Negative Control was not detected within 45 cycles.



The results of the Internal Control assay are shown above: each reaction hole was detected normally.

 

2) The experimental results for the Positive Control group are as follows:



The results of porcine-derived 16S rRNA assay are shown in the above figure: 1. Positive Control can obtain good amplification;



The results of eukaryotic 18S rRNA assay are shown above: 1. Negative Control was not detected within 45 cycles.



Internal Control The results are shown above: the reaction holes are detected normally.