RE-RRS of microbiome: sheep rumen
DNA extracted using a combined bead-beating, phenol and column purification protocol using the QIAquick 96 PCR purification kit. After digestion of DNA using the restriction enzyme PstI, barcodes were ligated to link sequences to samples, and samples were grouped in libraries consisting of four 96-well plates and PCR amplified. Amplified sequences between 193 and 318 bp were selected using a Pippin Prep and each library was run on an Illumina Novaseq 6000 machine, generating 101 bp single end reads using version 4 chemistry. Sequenced reads were demultiplexed using GBSX with default settings except that mismatches in barcodes or the cut site were disallowed.