Demultiplex without barcode file

Dear All,

Concerning the demultiplex stage I have some questions about the comands input below:

qiime demux emp-single
–i-seqs emp-single-end-sequences.qza
–m-barcodes-file sample-metadata.tsv
–m-barcodes-column BarcodeSequence
–o-per-sample-sequences demux.qza
–o-error-correction-details demux-details.qza

Q1- Is there any method to demultiplex without associate to barcode (In Qiime 1.9.0 I used the –i folderpairedsequence –o outputfolder –demultiplexing-method sampleid-by-file? Is there any similar command like this one?

2- What kind of file is the .qza? Is it the folder with the paired sequences? Can I just put the folder with the paired sequences as input?

3- The barcodes file is just an excel one save in .tsv? Do we need to put the barcodes and the samples name? How to transform into .fastq files? I just have them in excell.

I would greatly appreciate for any clarification.


1 Like

Hi @AndreFC, you need to know what barcodes represent what sample in order to demultiplex. The barcodes do not need to be saved in their own seperate “barcode reads” fastq file. One option is to use q2-cutadapt to demux, check out this tutorial:

Keep us posted! :t_rex:

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