Errors when demultiplexing paired end sequences

@sandray1007 Just chime in here, I did some search on this forum and it seems similar questions have been asked and answered (e.g. [Demuxing golay barcodes?]). You have to specify --p-rev-comp-barcodes and --p-rev-comp-mapping-barcodes at the same time, IF you are using the current version of EMP protocol, in which the Golay barcode is on the forward primer.

As explained by thermokarst here Demuxing golay barcodes? - #8 by thermokarst, "With Golay error correction, the orientation matters 100% — the barcode sequences and the mapping sequence both need to be in the 5′ -> 3′ orientation in order for". What this suggests is that the Golay barcodes in the current version of EMP protocol is actually in the wrong orientation: they are the reverse complement to the actual golay barcodes. That is why you have to set --p-rev-comp-barcodes to let QIIME know that the barcode reads need to be reverse-complemented before error correction, and --p-rev-comp-mapping-barcodes to let QIIME know to reverse-complement the barcode in the mapping file before demultiplexing.

If my understanding is correct, then I would think the EMP protocol needs to put a notice that the Golay barcode is not actually the Golay barcode but its reverse complement.

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