16S rRNA sequencing of TF sediment in acidic common garden
We conducted a single DNA extraction for each sample using 0.25 g of sediment and the DNeasy PowerSoil Pro kit (Qiagen, Hilden, DE) according to manufacturer instructions. The extracted DNA was used as template for 16S rRNA paired-end gene sequencing. The V4 region of the 16S rRNA gene was amplified in a 15 microL PCR reaction with dual indexed primers: 515F (5-GTGCCAGCMGCCGCGGTAA-3) and 806R (5-GGACTACHVGGGTWTCTAAT- 3) (Kozick et al. 2013). Amplified DNA served as template for high-throughput paired-end (2 X 250 bp) sequencing on an Illumina MiSeq v2 Standard flow cell at the Michigan State University Research Technology Support Facility.