Journal: Veterinary and Animal Science

Article Title: Development of duplex crystal digital PCR (dPCR) assay for detection and differentiation of NDRV and MDRV

doi: 10.1016/j.vas.2025.100502

Figure Lengend Snippet: Sensitivity comparison of duplex crystal dPCR and duplex qPCR(A-B): digital PCR sensitivity test, lane 1-6 is the digital PCR sensitivity dynamic curve of pNDRV and pMDRV with plasmid concentration of 1.5 × 10 4 to 1.5 × 10 -1 , lane 7 is NTC; (C-D): Standard curves for duplex qPCR and duplex crystal dPCR, which were prepared by 10-fold serial dilutions of pNDRV and pMDRV standard plasmids from 1.5 × 10 4 copies /µl to 1.5 × 10 0 copies /µl.

Article Snippet: The volume is 25 μL, including 12.5 μL of PerfeCta Multiplex qPCR ToughMix UNG (2 ×) (Quanta Biosciences, Gaithersburg, MD, United States of America), 2.5 μl of Fluorescein Sodium Salt (1 μM) (Apexbio Biotechnology, Beijing, China), 1.0 μL of NDRV primers (25 pmol/μL), 0.3 μL of NDRV-P (25 pmol/μL), 1.2 μL of MDRV primers (25 pmol/μL), 0.4 μL of MDRV-P (25 pmol/μL), 2.5 μl of DNA/cDNA template, and 1.5 μl of RNase-free water.

Techniques: Comparison, Digital PCR, Plasmid Preparation, Concentration Assay