Metabarcoding of Petromyzon marinus gut
Sea lamprey (Petromyzon marinus) were fed on lake trout (Salvelinus namaycush) in controlled aquaria at 5C, 10C, and 15C, then subjected to fasting periods (030 days). Digestive tract contents were collected, and DNA was extracted using the gMax Mini Genomic DNA Kit (IBI Scientific). A ~140 bp region of the 12S rRNA gene was amplified via PCR with vertebrate-specific primers and a lamprey-specific blocking primer. Dual-indexed Illumina sequencing adapters were added, and libraries were purified, quantified, and sequenced on an Illumina MiSeq (2  150 bp) at Michigan State University. Reads were processed in mothur (v1.48.0), aligned to a Great Lakes fish 12S database, and filtered with a 10 read and 1% RRA detection threshold.