Catfish metatranscriptome
The four heart samples were homogenised using a TissueLyser (Qiagen), and RNA was then extracted using an QIAamp Viral RNA Mini Kit (Qiagen) via a QiaCube extraction machine (Qiagen) according to the manufacturers instructions. RNA samples were then stored at -80oC. The RNA was converted to cDNA using Superscript II Reverse Transcriptase (Invitrogen, USA) following the manufacturers guidelines using 8 l of RNA and 1 l random hexamer primers per reaction. DNase was not used, allowing for any extracted DNA to also be sequenced.The concentration of the RNA/DNA samples for four samples was determined using a BioDrop Lite v1.0.4 (Thermo Fisher Scientific) and 7.5 l of each sample was barcoded using 2.5 l of Fragmentation Mix RB01-04 from Oxford Nanopores Rapid Barcoding Sequencing kit (SQK-RBK004, Oxford Nanopore Technologies). Barcoded samples were then equimolar pooled and sequencing adapters were ligated to the DNA library using 4 l of Rapid Adapter reagent (RAP, Oxford Nanopore Technologies). The barcoded library was loaded onto a MinION flow cell Mk1 R9.4.1 (Oxford Nanopore Technologies) and run via MinKNOW software v.3.6.5 (Oxford Nanopore Technologies) without real-time base-calling for 68 hours on a MinION sequencer (Oxford Nanopore Technologies).