Amplicon sequencing of RCP-PCR constructs
DB-X and AD-Y RCP-PCRs were performed by the protocol described in Yachie et al. 2016. In brief, the first PCRs, Row-Column PCRs (RC-PCRs), for DB-X barcodes and AD-Y barcodes were performed with the primer sets encoding Row or Column barcodes. The RC-PCR products were pooled by each plate and subjected to plate PCR using custom indexed primers for Illumina library preparation. The RCP-PCR library was quantified by qPCR  and sequenced by MiSeq (Illumina, 2250 bp paired-end sequencing).