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Table 3 Primers used for specific PCR and direct sequencing, amplification conditions and temperature profiles.

From: Why do snails have hairs? A Bayesian inference of character evolution

Primer Sequence amplification conditions temperature profile
COI universal [43] 5'-GGTCAACAATCATAAAGATATTGG-3' 5'-TAAACTTCAGGGTGACCAAAAAATCA-3' total volume 25 μl with: 0.17 mM dNTPs 3 mM MgCl2 in 1 × PCR buffer 0.13 μM of each primer 1 unit Taq polymerase (Invitrogen) 1 cycle of 2.5 min at 94°C 40 cycle 30s at 90°C 1 min at 48°C 1 min at 72°C 1 cycle of 10 min at 72°C
16S universal [44] 5'-CGGCCGCCTGTTT ATCAAAAACAT-3' 5'-GGAGCTCCGGTTTGAACTCAGATC-3' total volume 15 μl with: 0.1 mM dNTPs 2.5 mM MgCl2 in 1 × PCR buffer 0.2 μM of each primer 0.5 unit Taq polymerase (Invitrogen) 1 cycle of 2.5 min at 90°C 10 cycles of 50s at 92°C 30s at 44°C 40s at 72°C 36 cycles of 30s at 92°C 40s at 48°C 40s at 72°C 1 cycle of 3 min at 72°C
ITS-1 mollusc specific [45] 5'-TAACAAGGTTTCCGTAGGTGAA-3' 5'GCTGCGTTCTTCATCGATGC-3' total volume 15 μl with: 0.3 mM dNTPs 2.5 mM MgCl2 in 1 × PCR buffer 0.18 μM of each primer 0.5 unit Taq polymerase (Invitrogen) 1 cycle of 3 min at 94°C 40 cycles of 30s at 92°C 30s at 52°C 1 min at 72°C 1 cycle of 5 min at 72°C