title: Nanopore seq of Peromyscus: fecal sample031
design: DNA was extracted from fecal samples using a QIAamp PowerFecal Pro kit (Qiagen). The Oxford Nanopore Technologies (ONT) Rapid 16S Barcoding Kit (SQK-16S024; utilizing Kit 9 chemistry) was used to prepare barcoded amplicon libraries for sequencing. In brief: the full-length bacterial 16S rRNA gene region (1.6 kb) was amplified via PCR using specific primers and between 20-40 ng of DNA template, a long-range master mix, and sample-specific barcode identifier. PCR products were purified and prepared for sequencing through a series of magnetic bead wash steps. Barcoded samples were pooled (pools of 21-24 samples) with ONT rapid sequencing adapter mixture into a final library for sequencing. Pooled libraries were sequenced on FLO-MIN106 MinION flow cells utilizing R9 sequencing chemistry, run for 24 hours using the ONT MinKNOW GUI (v4.3.20).