Table 1.
Oligonucleotides used for PCR amplification of viral and transgene sequences
| Primer | Sequence (5'-3') | Target sequence |
| EAB555/F | (5'-TACATCGGCCTTTGAGTCGCATGG-3') | EACMV DNA B |
| EAB555/R | (5'-CTTATTAACGCCTATATAAACACC-3') | EACMV DNA B |
| JSP001 | (5'-ATGTCGAAGCGACCAGGAGAT-3') | ACMV (AV1/CP) |
| JSP002 | (5'-TGTTTATTAATTGCCAATACT-3') | ACMV (AV1/CP) |
| UniF | (5'-KSGGGTCGACGTCATCAATGACGTTRTAC-3') | CMGs DNA A |
| UniR | (5'-AARGAATTCATKGGGGCCCARARRGACTGGC-3') | CMGs DNA A |
| MSVF | (5'-ATCCCTCCAAATTCCGACAC-3') | MSV |
| MSVR | (5'-TCCATGTACAAAGCTCCTCT-3') | MSV |
| C1F | (5'-GCAGATCTATGCCTCGTTTATTTAAAATATATGC-3') | TYLCV |
| C1R | (5'-GCGGTACCTTACGCCTTATTGGTTTCTTCTTGGC-3') | TYLCV |
| TMVF | (5'-GCGGTGGCGGCCGATCCATGGAACTTACAG-3') | TMV |
| TMVR | (5'-GATTCGAACCCCTCGCTTTAT-3') | TMV |
| POT1 | (5'-gacgaattcTGYGAYGCBGATGGYTC-3') | TEV & PVY |
| POT2 | (3'-ACCACRTADCTBTTAcctaggtcag-5') | TEV & PVY |
| AC1F | (5'-ATGAGAACTCCTCGTTTTAGAA-3') | ACMV-Kenya AC1 |
| AC1R | (5'-ATGAGAACTCCT CGTTTTAGAA-3') | ACMV-Kenya AC1 |
| MP141 | (5'-ATGATTGAACAAGATGGATTGCAC-3') | NptII coding sequence |
| MP142 | (5'-TCAGAAGAACTCGTCAAGAAGGCG-3') | NptII coding sequence |
EACMV – East African cassava mosaic virus; ACMV – African cassava mosaic virus; MSV – maize streak virus; TYLCV – tomato yellow leaf curl virus; TMV – tobacco mosaic virus; TEV – tobacco etch virus; PVY – potato virus Y
EAB555/F &EAB555/R: PCR conditions consisted of 30 cycles of 94°C for 1 min, 58°C for 1 min. and 72°C for 2 mins.
JSP001 &JSP002: PCR conditions consisted of 30 cycles of 94°C for 1 min, 45°C for 1 min. and 72°C for 2 mins.
Uni F &Uni R: K = G + T, R = A + G, S = G + C. PCR conditions consisted of 30 cycles of 94°C for 1 min., 58°C for 1 min. and 72°C for 2 min.
MSV F &MSVR : PCR conditions consisted of 30 cycles of 94°C for 1 min, 59°C for 1 min and 72°C for 2 mins.
C1F &C2R: PCR conditions consisted of 94°C for 10 mins followed by 35 cycles of 94°C for 30 secs., 60°C for 1 min and 72°C for 1.5 mins followed and extension of 7 mins at 72°C.
AC1F &AC1R : PCR conditions consisted 94°C of 5 mins followed by 35 cycles of 94°C for 30 secs, 58°C for 1 min. and 72°C for 1 mins followed by extension of 10 mins at 72°C.
MP141 &MP142: PCR conditions consisted 5 mins. at 94°C followed by 35 cycles of 94°C for 30 secs, 58°C for 1 min. and 72°C for 1 mins. followed by extension time of 10 mins. at 72°C.