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We describe a method using technical replicates that are included in each run to determine and apply an appropriate adjustment per batch without manual intervention. A batch adjustment procedure is required to reduce variability across batches and to facilitate direct comparison of runs performed across multiple barcodes run over weeks, months, or years. However, with only 20 samples per barcode, multiple barcode sets (batches) are required to address questions in robustly powered study designs. A barcoding approach allows for 20 unique samples to be pooled and processed together in one tube, reducing the intra-barcode technical variability.
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