16s marker sequencing of S151_Cad_FvH6
The v4 hypervariable regions of archaeal and bacterial SSU rRNA genes were amplified using degenerate primers. The primers were 515F GTpad-GYCAGCMGCCGCGGTAA and 806rB GGpad-ACTACNVGGGTWTCTAAT. Amplification was done with fusion primers containing the 16S  sequences (including 2 nt pad) fused to Illumina adapters. The forward primers contain a 5 nt multiplexing barcode and the reverse a 6 nt index. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 10 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (ThermoFisher), 3 mM MgSO4, 0.2 uM of each primer and peptide nucleic acids to block host and plastid amplification. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair.