PacBio sequence run 1 of pooled PCR amplicons from 14 different R genes in Virola surinamensis DNA: Seedling root from seed collected under mother tree 78 and grown in soil containing a soil microbial inoculum from beneath mature mother tree 81
Amplicons of 14 selected R genes were produced by first round PCR using Phusion Hot Start II High Fidelity Polymerase and gene specific primers with the 3-prime end attached to the PacBio universal adapter sequences U1 or U2.  A 5 microliter aliquot of PCR product for each gene amplicon was pooled by individual plant sample and cleaned using solid phase reversible immobilization magnetic beads.  These sample amplicon pools (one pool per well of a 96-well plate) were submitted to the Penn State Genomics Core Facility for second round PCR with Kapa HiFi HotStart PCR Kit (KAPA Biosystems) to add a unique PacBio barcode to each sample amplicon pool.  After purification and molarity normalization barcoded amplicons were pooled and processed into SMRTbell Templates.  Circular consensus sequencing (CCS) of these Templates was on a PacBio Sequel System using Sequel chemistry 2.1 for plate one (sequencing run 1).  Movie time was 10 h.  After barcode parsing and CCS generation specifying minimum 10 passes and minimum predicted accuracy of 90% as the criteria for a CCS read, individual amplicons within a sample amplicon pool were deduced by alignment with a reference set of 14 "consensus" sequences (provided file Amplicons_14_WithoutPrimers.fa)  that derived from a prior transcriptome project.