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Review
. 2021 Jan;41(1):85-94.
doi: 10.5851/kosfa.2020.e80. Epub 2021 Jan 1.

Comparison of Seven Commercial TaqMan Master Mixes and Two Real-Time PCR Platforms Regarding the Rapid Detection of Porcine DNA

Soo Ji Kang  1 , Chan Song Jang  1 , Ji Min Son  1 , Kwang Won Hong  1
Affiliations
Review

Comparison of Seven Commercial TaqMan Master Mixes and Two Real-Time PCR Platforms Regarding the Rapid Detection of Porcine DNA

Soo Ji Kang et al. Food Sci Anim Resour. 2021 Jan.

Abstract

A pig-specific real-time PCR assay based on the mitochondrial ND5 gene was developed to detect porcine material in food and other products. To optimize the performance of assay, seven commercial TaqMan master mixes and two real-time PCR platforms (Applied Biosystems StepOnePlus and Bio-rad CFX Connect) were used to evaluate the limit of detection (LOD) as well as the PCR efficiency and specificity. The LODs and PCR efficiencies for the seven master mixes on two platforms were 0.5-5 pg/reaction and 84.96%-108.80%, respectively. Additionally, non-specific amplifications of DNA from other animal samples (human, dog, cow, and chicken) were observed for four master mixes. These results imply that the sensitivity and specificity of a real-time PCR assay may vary depending on master mix and platform used. The best combination of master mix and real-time PCR platform can accurately detect 0.5 pg porcine DNA, with a PCR efficiency of 100.49%.

Keywords: master mix; porcine DNA; real-time PCR; species identification.

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Conflict of interest statement

The authors declare no potential conflicts of interest.

Figures

Fig. 1.
Fig. 1.. Standard curves for seven master mixes (a–g) tested on the StepOnePlus and CFX Connect platforms.
The real-time PCR assay was completed in triplicate using 10-fold serial dilutions of porcine DNA. Error bars are not shown because the symbol is larger than the error bar.
See this image and copyright information in PMC

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