High-throughput accuracy for multiplex amplicon sequencing

Solution to the new problem of mistagging leading to intractable cross-conmtamination as a result of sample-barcode switching. Different mulitplexing experimental desings are sequenced as different sequencing Illumina TruSeq libraries to detect, measure and solve the mistagging issue. Two additional libraries are sequenced to prove the accuracy of a new filter taking advantage of mistagging patterns and of the incorporazion of PCR replicates. Based on the refined experimentsl planning and computational analyses of the distribution od random and non-random errors, we propose a solution toward high-throughput accuracy in multiplex amplicon sequencing experiments for diversity analysis.