Table 3. Primers used for RTqPCR experiments.
| primer name | target gene | nucleotide sequence | Tm (°C) | amplicon size (nt) | PCR efficiency in protoplast RNA samples | PCR efficiency in plant RNA samples |
|---|---|---|---|---|---|---|
| TYMV 150–30 TYMV 150–31 |
TYMV genomic RNA |
CAACTGGTCTTATACCCACTTCCG CGGAATGGACCATGGGTAGG |
65.2 62.5 |
191 nt | 94.1% | 100.7% |
| EF1 F2 EF1 R1 |
EF1α (At5g60390) |
TGGTAACGGTTACGCCCCA ACCAGCGTCACCATTCTTCAAA |
59.5 60.3 |
142 nt | 94.5% | 97.8% |
| PDF2 F5 PDF5 R5 |
PDF2 (At1g13320) |
AGTGAGAACAATGACGATGACG GCATACTCAACCCCTCCTACATAC |
60.3 65.2 |
92 nt | 91.6% | not done |
| 18S F 18S R |
18S rRNA |
GGGAAACTTACCAGGTCCAGACATAG CCACGTAGCTAGTTAGCAGGCTGAG |
67.9 69.1 |
159 nt | not done | 102.9% |