title: 18S amplicons from E. superba gut contents
design: Nested PCR was used to amplify 18S from krill gut contents. In the first PCR, krill stomach DNA extracts were amplified with universal 18S primers, while krill s own DNA was blocked with a krill specific PNA probe. These reactions included 1 x GoTaq Green Master Mix (Promega), 0.5  M each forward and reverse primers (forward: 5 -GGGCATCACAGACCTG-3  reverse: 5 -GGCTYAATTTGACTCAACRCG-3 , modified from Gast et al. 2004 as per Cleary et al. 2015), 20  M krill-specific PNA (from 100  M stock re-suspended in 2.5% Trifluoroacetic acid, 5 -CGTCGGGTTGTCTTG-3  (Cleary et al. 2012)), and 20% by volume stomach DNA extract. In the second PCR, reactions contained 1 x GoTaq Green Master Mix, 0.2  M each forward and reverse primers, and 20% by volume amplicons from the first round of PCR. Illumina primers were reversed, which is to say the read1 adaptor (traditionally forward) was put on the reverse primer, and the read2 adaptor on the forward primer (Read1 primer: CGTCGGCAGCGTCAGATGTGTATAAGAGACAGGGCTYAATTTGACTAACRCG; Read2 primer: GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGGGCATCACAGACCTG).
library_name: MCC56