Table 2. Cyclopia specific primers designed for testing HRM haplotype discrimination.
Primers used to screen haplotype variation in wild C. subternata populations are indicated in bold. Primer details provided include; non-coding cpDNA region the primers are located in, as well as each primers’ annealing temperature (Tm) in degrees Celcius, GC content, and sequence motif.
| Region | Primer | Direction | Tm (°C) | GC (%) | Sequence (5′→3′) |
|---|---|---|---|---|---|
| trnG intron | MLT_C1 | F | 57.3 | 43.5 | ACTCCTCTTCTATTCATGGGGA |
| MLT_C2 | R | 58.0 | 50.0 | ||
| MLT_C3 | F | 61.8 | 40.9 | TCAACGAACGATTCGAGGAATA | |
| MLT_C4 | R | 61.1 | 45.5 | TGCTTCAATCTCTCCTACCCAA | |
| pctL-psbE intergenic spacer | MLT_M1 | F | 58.0 | 43.5 | TGTCGAGAACCCTTATACTCTCA |
| MLT_M2 | R | 58.7 | 47.6 | TACCAAGGGTGTCTTTCGAGT | |
| atpI-atpH intergenic spacer | MLT_S1 | F | 64.3 | 50 | ATTACAGATGAAACGGAAGGGC |
| MLT_S2 | R | 61.5 | 45.5 | TGGGGGTTTCAAAGCAAAGG | |
| MLT_S3 | F | 61.5 | 45.5 | CCTTTGCTTTGAAACCCCCA | |
| MLT_S4 | R | 66.4 | 36.5 | TTCCCGTTTCATTCATTCACATTCA | |
| ndhA intron | MLT_U1 | F | 59.1 | 40.0 | AGGTACTTCTGAATTGATCTCATCC |
| MLT_U2 | R | 62.2 | 52.4 | GCAGTACTCCCCACAATTCCA | |
| rpl32-trnL intergenic spacer | MLT_V1 | F | 59.9 | 60.0 | CTCCTTCCCTAAGAGCAGCG |
| MLT_V2 | R | 59.2 | 40.0 | GTTGGAATAATCTGAATTAGCCGGA |