【Product Description】
This product is a freeze-dried real-time PCR detection reagent for pig (Sus scrofa) ingredient in food designed according to method in the China National Standard GB/T38164-2019. It is a mastermix with all ingredients necessary for real-time PCR amplification of the Sus scrofa ATP8 gene. It is pre-dispensed in individual PCR tubes, and can be used directly in a PCR machine after adding sample DNA. It does not require cold chain for storage and transportation, thus reducing risk and cost significantly. It is particularly suitable for on-site fast detection of pig ingredient in food.
spec. | Description | QTY |
FS-AI-03 | Freeze-Dried Real-Time PCR Detection Reagent for Pig Ingredie | 48T |
FS-AI-03PC | pig ingredient positive control | 1 tube |
EP-CM-10 | Resealable bag | 1 bag |
【Storage and Shelf Life】
Store at ambient temperature (4-35℃). Stable for 12 months from date of manufacturing. This product is packed in PCR strips in vacuum with desiccant in an aluminum bag. Keep the reagent dry and avoid exposure to light before use. Please store the unused reagent in the provided re-sealable bag with desiccant, and use within a week after breaking the vacuum.
Note: The reagent pallets will shrink if dampened by moisture. Discard reagents with significant shrank pallets, or tested with positive control before used for sample testing.
【Device or Reagents Not Included】
1) Pipettes and filtered tips.
2) Optical tube caps for real-time PCR.
3) Nuclease-free water.
4) Nucleic acid extraction kit.
【Suitable Specimen】
Meat/meat products, animal organs, milk, animal feed, etc.
【Instruction for Use】
1. DNA Extraction
Extract DNA using an appropriate extraction kit for different specimens. Use 100μl elution buffer (TE/ nuclease-free H2O) to elute DNA at the final step of the extraction protocol. Use the purified DNA immediately or store at -20℃ for later use.
2. Preparation of Positive control
The positive control in this kit is a freeze-dried plasmid DNA fragment of the pig ATP8 gene (colorless pallet). It is stable for up to 12 months before re-hydration. Re-hydrate the positive control with 250μl of nuclease-free H2O, set on ice for 10 minutes, vortex at low speed for to mix well. Use immediately or store at -20℃ for later use.
3.Preparation of Real time PCR mix
1) Open the vacuum package containing the reagent strips, and check the reagent pallet is on the bottom of the tube.
2) Open and discard the strip caps (not suitable for Real Time PCR). If the tubes are not fit for the real-time machine, transfer the reagent to suitable PCR tubes for testing.
3) Prepare the Real-Time PCR MasterMix on ice as following:
Components | vol. |
Real-Time PCR Detection Reagent for Pig Ingredient | 1 tube(2ul) |
Sample DNA/Positive Control/No Template Control* | 23 ul |
Total | 25 ul |
*Use nuclease-free H2O as no template control.
4) Cap the PCR using caps (strips) suitable for real time PCR.
5) Vortex at low speed for 10-20 seconds to mix well, and centrifuge at 3000rpmfor 20 seconds, and put in a real time PCR machine for testing
4. Real Time PCR set up
1) Set up the reaction for sample/positive control/no template control according to instructions of the real-time PCR machine.
2) Set reaction volume as 25μl.
3) Set report fluorescent dye as FAM, quencher dye as TAMRA, reference dye as NONE.
4) Set temperature profile as the following and collect
fluorescence at 60℃.

5. Result Interpretation
