Problem Demultiplexing Sequences (Solved)

Edit: Qiime took an exceptionally long time with the last block of command lines, but the Demux worked at last!

Hi. I am trying to Demux on Qiime2-2019.7, but I am running into several problems. I have been following the tutorials and discussions in this forum. However, these are my commands and error messages:

$ qiime demux emp-single --i-seqs emp-single-end-sequences.qza --m-barcodes-file 54587_mapping_file.txt --m-barcodes-column BarcodeSequence --o-per-sample-sequences demux.qza --o-error-correction-details demux-details.qza

Plugin error from demux:

  • No sequences were mapped to samples. Check that your barcodes are in the correct orientation (see the rev_comp_barcodes and/or rev_comp_mapping_barcodes options).*

Debug info has been saved to /tmp/qiime2-q2cli-err-wfvw1wl6.log

I have also tried the same with:

–p-no-golay-error-correction

But received the same the same error message.

As well as:

$ qiime demux emp-single --i-seqs emp-single-end-sequences.qza --m-barcodes-file 54587_mapping_file.txt --m-barcodes-column BarcodeSequence --o-per-sample-sequences demux.qza --p-golay-error-correction --o-error-correction-details data_golay_errors --p-rev-comp-barcodes --p-rev-comp-mapping-barcodes

But Qiime seems to be stuck with no progress for this case.

What could I be doing wrong here? Cheers!

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–p-rev-comp-barcodes --p-rev-comp-mapping-barcodes

What orientation are your barcodes are in? forward or reverse primers?

I would try to remove one or the other:
–p-rev-comp-barcodes
–p-rev-comp-mapping-barcodes

edit: nm I just read your messages - yes, it should generate a file that’s 3-6 gigabytes. Ben

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