SNP | Primer sequence | PCR conditions | Amplicone Size (bp) | RFLP | |||
---|---|---|---|---|---|---|---|
Restriction enzymes | Incubation conditions | Product after digestion (bp) | |||||
Ancestral | Variant | ||||||
rs3917655 | 5’TGTCCACTTTGACCCTCCCA3’ 5’AGGGCAGAAAAGGAAACTATGTG3’ | Initial denaturation at 95 °C (7 min) 30 s at 95 °C 30 s at 58 °C 30 s at 72 °C Final elongation at 72 °C for 7 min | 405 | PvuII | At 37 °C for 2 h | 249 | 405 |
156 | |||||||
rs3917657 | 5’ATCTTCTGGGACTGATCTGGA3’ 5’CCTGCCTGGTTCCTCCATAG3’ | Initial denaturation at 95 °C (7 min) 30 s at 95 °C 30 s at 60 °C 30 s at 72 °C Final elongation at 72 °C for 7 min | 516 | TfiI | At 65 °C for 2 h | 265 | 265 |
251 | 199 52 | ||||||
rs3917739 | 5’AAAGCCCAGAGCAAAGAGGTAGT3’ 5’CCCTCCCTTCCCACCTTAACT3’ | Initial denaturation at 95 °C (7 min) 30 s at 95 °C 30 s at 60 °C 30 s at 72 °C Final elongation at 72 °C for 7 min | 546 | TfiI | At 65 °C for 2 h | 546 | 328 |
218 | |||||||
rs3917843 | 5’ATTACATGCAATGCCTGCCT3’ 5’GGGGCATACTGTCCCTTTTTGA3’ | Initial denaturation at 95 °C (7 min) 30 s at 95 °C 30 s at 59 °C 30 s at 72 °C Final elongation at 72 °C for 7 min | 578 | BsaWI | At 60 °C for 2 h | 329 | 578 |
249 | |||||||
rs2235302 | 5’GCCAACCTGTGAGGGTAGGAT3’ 5’ACCACTGTCCGCCTTATAAACT3’ | Initial denaturation at 95 °C (7 min) 30 s at 95 °C 30 s at 57 °C 30 s at 72 °C Final elongation at 72 °C for 7 min | 511 | EciI | At 37 °C for 2 h | 441 | 511 |
70 |