Introduction

Barcodes are used in NGS to tag samples before pooling. After sequencing, these barcodes are used to demultiplex the data, thereby assigning the reads to the originating sample.

The key aspect of a good set of barcodes is robustness against read errors. One read error should not be able to transform one barcode into another. This requirement can be met by selecting barcodes in such a way that the edit distance between any pair of barcodes is larger than one. An additional desired property is the ability to correct read errors. This can be done by increasing the minimal edit distance between barcodes to at least three. If one read error occurs, the sequenced barcode will have a distance of one to the original barcode and a minimum distance of two to any of the other barcodes. If the read error is high, the minimum edit distance should be increased to a higher (odd) number.

For some sequencers it is important that mononucleotide stretches in barcodes are below a minimum length. An additional filter can be used to remove these barcodes.