title: Raccoon oral gut microbiome - USA_F6
design: DNA was extracted from swabs using the NucleoSpin Tissue XS kit for DNA from cells and tissue (Macherey-Nagel) according to the manufacturer's protocol in the pre-PCR dedicated area. PCR was performed with modified indexed bacterial primers 16SF_ins0 (GTGCCAGCMGCCGCGGTAA), 16SF_ins3 (TGAGTGCCAGCMGCCGCGGTAA), 16SR_ins0 (GGACTACHVGGGTWTCTAATCC) and 16SR_ins3 (TGAGGACTACHVGGGTWTCTAATCC) (Galan et al., 2016). A second PCR was performed using indexed i5 and i7 primers enabling two step indexing. Per each plate we used a combination of eight unique forward and twelve unique reverse primers with 8-base indexes.