Table 1. PCR primer sets and amplification conditions of loci used in Experiments I (only primer sets that amplified fragments representing single locus polymorphism are shown) and II.
| Experiment | Locus | Primer sequence (5′-3′)/Source | Tm (°C)a | PCR cycles | Primer (p)b | MgCl2 (m)c | Species testedd | GenBank accession no.e |
|---|---|---|---|---|---|---|---|---|
| I | Gcin1 | F: TCAAATTATGTATCACAAATCTCA R: AAAGTAACCAGTCTCGGGCTGTCG | 51 | 25 | 1 | 2.5 | GC, GU, GM, CZ, OL | GQ129680 |
| I | Gcin2f | F: CTTTTTGGGCCTGATGCTTCTG R: GATCCCGAGGTGCTTATGGTATG | 58 | 25 | 2 | 2.0 | GC, GU, GM, CZ, OL | GQ129681 |
| I | Gcin3f | F: TTGGGCATGATGCTTCTGGAG R: GATCCCGATGTGCTTATGGTATG | 58 | 25 | 1 | 2.0 | GC, GU, GM, CZ, OL | GQ129682 |
| I | Gcin4 | F: CTAGCATGCTATAGTTTAGGAGAT R: CGCCGAGCGTATTTTTA | 48 | 25 | 2 | 3.0 | GC, GU, GM, CZ, OL | GQ129683 |
| I | Llit52 | F: GTATTGTTGTCTTGCACCACGAA R: ATCCAACGAATTTAGACACAACCA | 56 | 25 | 1 | 2.5 | LS, LL, LF, LO, LC | FJ480222 |
| I | Lsax12 | F: AAAATGCCGAAGCGACAGT R: TGCCGAAAACCAAACCA | 53 | 25 | 1 | 2.5 | LS, LL, LF, LO, LC | FJ794786 |
| II | GcinCvA | F: GAGTCATTGGTTTCGGAGAAGAA R: GCTCGATCTCATCAGGTCTGAAA | 58 | 25 | 2 | 2.5 | GC, GU, GM, CZ, OL, LS, LL, LF, LO, LC | GQ129736 |
a
Annealing temperature.
b
Primer concentration pmol.
c
Magnesium chloride concentration.
d
Species tested: CZ, Calliostoma zizyphinum; GC, Gibbula cineraria; GM, G. magus; GU, G. umbilicalis; LS, Littorina saxatilis; LC, L. compressa; LL, L. littorea; LF, L. fabalis; LO, L. obtusata; OL, Osilinus lineatus.
e
GenBank accession numbers for the cloned sequences from which the markers were derived.
f
Instances where a touchdown PCR profile was used.