Demultiplexing paired end read Problem

#import data
qiime tools import
–type ‘SampleData[PairedEndSequencesWithQuality]’
–input-path pe-64-manifest
–output-path paired-end-demux.qza
–source-format PairedEndFastqManifestPhred33

qiime demux emp-paired
–m-barcodes-file metadata_4samples.tsv
–m-barcodes-column BarcodeSequence
–i-seqs paired-end-demux.qza
–o-per-sample-sequences demux

qiime demux summarize \
  --i-data demux_4samples.qza \
  --o-visualization demux_4samples.qzv

#error message
Plugin error from demux:

Argument to parameter 'seqs' is not a subtype of EMPPairedEndSequences.

Debug info has been saved to /tmp/qiime2-q2cli-err-ya8xuuh2.log

I could not see the logfile, actually, Could you help me figure out what’s wrong?
Thank you.

Again, the error message

How to identify the
DATA2 trimming and denoising size?
I could not view the demux.qzv file,
If I coud view it, how to identify the trim & tranc size?
Thank you so much.
qiime dada2 denoise-paired
–i-demultiplexed-seqs paired-end-demux.qza
–p-trim-left-f 0???
–p-trim-left-r 0???
–p-trunc-len-f 0???
–p-trunc-len-r 0???
–o-representative-sequences rep-seqs-dada2.qza
–o-table table-dada2.qza
–o-denoising-stats stats-dada2.qza

I put numbers randomly, however, I run successfully without any error, but without any outputfile.
Could you help me please?
THank you.

My story still going on.
I don't know how could I make this step happen....change sever to mac book maybe help me this moment.
However, the table.qzv SEEMs not qualified compare with the results discussed on qiime2 forum.

Could you please let me know, What happened?
Thank you.


for the --p-sampling-depth
according to table.qzv ? frequency? or sequence account?

Thank you so much,

The alpha compare figure looks weird.

and I think BMI , BarcodeSequence, gender, sample-type, sequencing-run all listed in the metadata, why I got that information?

metadata listed below.

Thank you so much.

Hey @doudou2047 - looks like you made it through your initial problems if you made it this far, so I won’t address those posts.

This is because you only have 2 samples in each group.

As the warning states, those columns were ignored because they were either all unique (BarcodeSequence) or all the same value (sequencing-run, sample-type, gender).

Hi,thank you so much for your reply.
Could you please let me know the classifier based on the Greengenes? Is there any other options? or update?
I just directly use this one? or ?
The bar-figure such like that?

Thank you,

Seems the file upload unsuccessfully.
This is the part of taxa-bar-plots.qzv
taxa-bar-plots.qzv (334.2 KB)

Even I went through this step, I still confusing about the trim size.
The numbers I just copy from one example from qiime2 forum.
Could you let me know your suggestion?
Thank you.

Have you read the DADA2 Documentation? This will be the best way for you to learn how this plugin works.

I am going to lock this thread now, we request that users keep it limited to one topic per thread - if you have any additional questions, please open them in new threads, individually. Thanks!