Metabarcoding of Gallotia galloti gut microbiome, 16SV34
Samples were amplified and sequenced by Novogene (UK). Polymerase chain reaction (PCR) was conducted using a Phusion High fidelity PCR master mix (New England Biolabs, Cat# M0531S). The 16SV34 region was amplified using a specific forward and reverse primer (F: 5CCTAYGGGRBGCASCAG3, R:5GGACTACNNGGGTATCTAAT3) (Youngseob et al. 2005) and connecting barcodes. The amplified PCR was electrophoresed in 2% agarose gels and proper sized products were selected. Equal quantities of the selected PCR products were pooled, subjected to end repair, A-tailing and subsequently ligated with Illumina adapters. Samples were sequenced on an Illumina platform (Illumina NovaSeq 6000) which generated raw reads of 250 bp