Fragment | Primer | Nucleotide position | Sequence | Product size (bp) | MgCl2 conc (mmol/l) | Annealing temp (°C) | Restriction enzyme before SSCP (fragment sizes (bp)) | ||||
1 | SFA1(+) | 3-191-150 | M13 (-21)- GCGGGAGACCAGAGAGC | 301 | 1.5 | 64 | |||||
SFA1(−) | 267-251 | M13 rev-CAGCGGGGGATACAAGG | BstNI (118+183) | ||||||||
2 | SFA2(+) | 11-30† | M13 (-21)- ACAGTCCCAGCCTCCCTACT | 362 | 1.5 | 60 | |||||
SFA2(−) | 336-319 | M13 rev-ATGGGAGACGTGGCAGAG | HaeII (142+220) | ||||||||
3 | SFA3(+) | 79-95‡ | M13 (-21)- GGGCCATGGGAGAACAG | 306 | 1.5 | 62 | |||||
SFA3(−) | 348-330 | M13 rev- CGTGCCTTGGTACAAGGTG | ApaLI (129+177) | ||||||||
4 | SFA4(+) | 426-442‡ | M13 (-21)- CGAGAACCCACAGTGCG | 338 | 1.5 | 64 | |||||
SFA4(−) | 727-711 | M13 rev- GTGCTCTGGTACCGGCC | BsaAI (138+200) | ||||||||
5 | SFA5(+) | 27-43§ | M13 (-21)- CCTTCCTGTGCCACGTG | 354 | 1.0 | 62 | |||||
SFA5(−) | 344-323 | M13 rev- TCTTTCTTCATCATCTAGGGCC | RsaI (139+215) | ||||||||
6 | SFA6(+) | 493-511§ | M13 (-21)- TGTTCTAAGCCTGGCTCCC | 262 | 1.0 | 62 | |||||
SFA6(−) | 718-701 | M13 rev- CTGCCACACTGGACCCTC | StyI (130+132) | ||||||||
7 | SFA7(+) | 40-56¶ | M13 (-21)- GGATGCAGCAGCAGGTG | 422 | 1.5 | 64 | |||||
SFA7(−) | 425-406 | M13 rev- AGGTAATGGGTGTGGAGCAG | RsaI (219+203) | ||||||||
8a | SFA8a(+) | 1286-1304¶ | M13 (-21)- TTGGATTGGAGAAGGGAGC | 493 | 1.5 | 66 | |||||
SFA8a(−) | 1742-1721 | M13 rev- CCGGTAGTAGTAATGACGGAGG | HaeII (230+263) | ||||||||
8b | SFA8b(+) | 1632-1651¶ | M13 (-21)- CCCATCGACCAGGACTTCTA | 407 | 1.0 | 62 | |||||
SFA8b(−) | 2002-1984 | M13 rev- GGATGTGTCTGGGACATGC | SacI (143+264) |
↵1-150 † ‡ § ¶Positions of primers are numbered according to Genbank database entry u60107, u60108, u60109, u60110, and u60111, respectively.7