title: 16S/ITS rRNA of buffelgrass Mexico project
design: To assess bacterial/archaeal communities, we performed PCR amplification of the V4 hypervariable region of 16S rRNA gene using the primer sets 515-F (GTGCCAGCMGCCGCGGTAA) and 806-R (GGACTACHVGGGTWTCTAAT) (Walters et al., 2015). For fungal communities, PCR amplification targeted the first internal transcriber region (ITS1) using the primers ITS1-F (CTTGGTCATTTAGAGGAAGTAA) and ITS2 (GCTGCGTTCTTCATCGATGC) (Walters et al., 2015). The primers included Illumina adapters, and the reverse primer pairs included unique error-correcting 12-base-pair barcode specific to each sample for demultiplexing. Each sample underwent triplicate PCR reactions in a 40 ml volume, consisting of 3 ml of extracted DNA, 3 ml of each primer, 20 ml of MyFi PCR Mix (Bioline, Taunton, MA, USA), and 11 ml of water.