5930_1
16S rRNA gene library of Marum Crater fumarole
DNA was extracted from 0.3 g of sample material using the PowerSoil DNA Isolation Kit (MoBio, Carlsbad, CA) with the supplied protocol. Following PCR validation (including positive and negative controls) with the 27F and 1492R primers (Weisburg et al. 1991), target regions were amplified using oligonucleotide primers 515F (5-GTGYCAGCMGCCGCGGTAA-3) (Parada, Needham and Fuhrman 2016) and 806R (5-GGACTACNVGGGTWTCTAAT-3) (Apprill et al. 2015), which were modified from the original 515F-806R primer pair (Caporaso et al. 2011). Purified amplicons were diluted to equal concentrations and were sequenced (2 x 250 bp) on an Illumina (San Diego, CA, USA) MiSeq sequencer at RTL Genomics (Lubbock, TX, USA).
5930_1
SUB6916237
5930_1