Viral DNA extraction was conducted using the Viral Gene-spin Viral DNA/RNA Extraction Kit (iNtRON Biotechnology, Seongnam, Korea) from tissue samples following manufacturer’s protocol. PCR analysis was done with 1× AccuPower PCR Master Mix (Bioneer, Daejeon, Korea) and TYLCV-specific primer set, whose target is TYLCV partial sequence for intergenic region (IR) and V2 gene (5′-GATGGCCGCGCCTTTTCCTTTTATGTGG-3′, 5′-GCTGCTGTATGGGCTGTCGAAGTTCAG-3′) (Kil et al., 2016 (link)). Amplification was carried out under the following conditions: preheating at 94°C (3 min), then 35 cycles of 30 s at 94°C, 30 s at 58°C and 1 min at 72°C, followed by a final extension at 72°C (5 min). PCR products were visualized in 1% agarose gel electrophoresis.
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Viral DNA Extraction and PCR Detection of TYLCV
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Agarose gel electrophoresis
Gene
Intergenic region
Intergenic sequence
Intron
Primer
Tissue
Viral dna
Viral gene
Viral rna
Corresponding Organization :
Other organizations : Andong National University, Sungkyunkwan University, Kyungpook National University, Rural Development Administration, National Academy of Agricultural Science
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Variable analysis
independent variables
- Viral DNA extraction method (Viral Gene-spin Viral DNA/RNA Extraction Kit)
- Presence/detection of TYLCV partial sequence for intergenic region (IR) and V2 gene
- Tissue samples
- AccuPower PCR Master Mix
- TYLCV-specific primer set
- PCR amplification conditions (preheating at 94°C for 3 min, 35 cycles of 30 s at 94°C, 30 s at 58°C and 1 min at 72°C, followed by a final extension at 72°C for 5 min)
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