Table 1.
Sequences and annealing temperature of the primers used for the genotyping by PCR-RFLP assays (A) and for preparation of the FISH probes covering the casein genes cluster (B). All primers were designed on wild feral camel genome sequence available in gene bank (EMBL acc. no. AGVR01039100.1), and multiple alignment confirmed 100% similarity in the other camelids.
| SNP (A) | GenBank ID | Primers | Annealing temperature (°C) | Size (bp) | Genotyping | |
|---|---|---|---|---|---|---|
| CSN1S1 | JF429138 | Forward: | 5’-TGAACCAGACAGCATAGAG-3’ | 58.5 | 930 | SmlI |
| c.150G > T | Reverse: | 5’-CTAAACTGAATGGGTGAAAC-3’ | ||||
| CSN2 | HG969421 | Forward: | 5’-GTTTCTCCATTACAGCATC-3’ | 60.0 | 659 | HphI |
| g.2126A > G | Reverse: | 5’-TCAAATCTATACAGGCACTT-3’ | ||||
| CSN3 | HE863813 | Forward: | 5’-CACAAAGATGACTCTGCTATCG-3’ | 62.0 | 488 | AluI |
| g.1029T > C | Reverse: | 5’-GCCCTCCACATATGTCTG-3’ | ||||
| Probe (B) | Gene | Primers | Annealing temperature (°C) | Size (bp) | Position | |
| 1 | CSN1S1 | Forward | 5’-GTACCCAGAAGTCTTTCAA-3’ | 59.5 | 913 | Exon 3 |
| Reverse | 5’-CACTGCTAACTCAAGAATCT-3’ | Exon 5 | ||||
| 2 | CSN2 | Forward | 5’-TTCACTTCTTTTCCTCCAC-3’ | 62.3 | 2433 | Exon 1 |
| Reverse | 5’-CCATTGTATTTGTGCAATATTA-3’ | Intron 1 | ||||
| 3 | CSN2 | Forward | 5’-GATGAACAGCAGGATAAAATC-3’ | 56.0 | 657 | Exon 7 |
| Reverse | 5’-ATCACTGATCTGAACTAT-3’ | Intron 7 | ||||
| 4 | CSN1S2 | Forward | 5’-AGCTGTAAGGAACATAAAGG-3’ | 60.5 | 1493 | Exon 7 |
| Reverse | 5’-TGTGGGGACTTCAGCTG-3’ | Exon 8 | ||||
| 5 | CSN3 | Forward | 5’-TGCAGAGGTGCAAAACCA -3’ | 61.5 | 1337 | Exon 4 |
| Reverse | 5’-GCTAGTCTGTGTTGGTAGTAA-3’ | Exon 5 | ||||