18S amplicon sequencing on Heterotermes tenuis protist hindgut community: 11.0228 S 40.5111 W
DNA was extracted from the whole gut contents of live termites or the macerated abdomens of termites preserved in ethanol or RNAlater. Each DNA template was acquired from a single worker individual. Amplicon libraries were generated using a two step PCR protocol. The first PCR used gene specific primers with Illumina adaptor sequences at their 5 ends: nexF_ParaV45F 5TCG TCG GCA GCG TCA GAT GTG TAT AAG AGA CAG 3 and nexF_ParaV45R 5GTC TCG TGG GCT CGG AGA TGT GTA TAA GAG ACA G 3. The second PCR attached indexing barcodes to both ends of each amplicon. PCR products were purified using Ampure  magnetic beads in combination with the Beckman Biomek NXp robot and quantified using Qubit broad range dsDNA fluorescent dye (Invitrogen) on a Biotek HT1 Plate Reader. After quantification, samples were pooled at approximately equivalent concentrations and sequenced on the Illumina MiSeq platform.