Paleo-faeces 3
All pre-PCR amplication steps were carried out in aDNA laboratories at the Centre for GeoGenetics applying established aDNA protocols. Extractions for shotgun metagenome sequencing were carried out using a phenol-chloroform- and kit-based extraction protocol optimized for ancient sedimentary and faecal samples. This entailed dissolving a total of ~16 g of palaeo-faecal matter in 40 ml digestion buffer. Libraries were constructed using the NEBNext DNA Library Prep Master Mix for 454 (E6070) and sequenced on an Illumina HiSeq 2500 platform at the Danish National High-Throughput DNA Sequencing Centre.