Illumina MV1 sequencing of seawater eDNA: Lindblad Expeditions Southern Ocean Sample 008C January/February 2022 Cruise
PCR products were amplified using the following primer sets: 12s MarVer1 (F): NeXT Fusion: 5TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCGTGCCAGCCACCGCG3 and 12s MarVer1 (R): NeXT Fusion: 5GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGGGTATCTAATCCYAGTTTG3. PCR products were generated using the following mix: 12.5L Q5 2X MasterMix (NEB), 9.5L nuclease-free water, 1uL 10M primer mix (0.4uM final), and 2L of 1:10 diluted DNA template. Samples were cycled using the following profile: initial denaturation at 95C for 3min, followed by 40 cycles of 95C for 30s, 49.5C for 30s, and 65C for 45s, followed by a final extension of 65C for 5min. After 0.8x bead cleanup, samples were then subjected to 8 cycles of indexing PCR using Nextera XT barcode cycling conditions before sequencing.