Hi @cherman2
I tried suggestions. Still, there is an issue. So my sequences imported to and able to create paired-end.qza. But, later command failed.
(qiime2-metagenome-2024.5) bhagwanrekadwad@MicrobeAI ~ % cd Desktop/shotgun
(qiime2-metagenome-2024.5) bhagwanrekadwad@MicrobeAI shotgun % qiime demux summarize
--i-data paired-end-demux-shotgun.qza
--o-visualization demux.qzv
Saved Visualization to: demux.qzv
(qiime2-metagenome-2024.5) bhagwanrekadwad@MicrobeAI shotgun % qiime demux emp-paired
--m-barcodes-file sample-metadata.tsv
--m-barcodes-column sample-id
--p-rev-comp-mapping-barcodes
--i-seqs paired-end-demux-shotgun.qza
--o-per-sample-sequences demux-full.qza
--o-error-correction-details demux-details.qza
Usage: qiime demux emp-paired [OPTIONS]
Demultiplex paired-end sequence data (i.e., map barcode reads to sample ids)
for data generated with the Earth Microbiome Project (EMP) amplicon
sequencing protocol. Details about this protocol can be found at
Protocols and Standards : earthmicrobiome
Inputs:
--i-seqs ARTIFACT EMPPairedEndSequences
The paired-end sequences to be demultiplexed.
[required]
Parameters:
--m-barcodes-file METADATA
--m-barcodes-column COLUMN MetadataColumn[Categorical]
The sample metadata column containing the per-sample
barcodes. [required]
--p-golay-error-correction / --p-no-golay-error-correction
Perform 12nt Golay error correction on the barcode
reads. [default: True]
--p-rev-comp-barcodes / --p-no-rev-comp-barcodes
If provided, the barcode sequence reads will be
reverse complemented prior to demultiplexing.
[default: False]
--p-rev-comp-mapping-barcodes / --p-no-rev-comp-mapping-barcodes
If provided, the barcode sequences in the sample
metadata will be reverse complemented prior to
demultiplexing. [default: False]
--p-ignore-description-mismatch / --p-no-ignore-description-mismatch
If enabled, ignore mismatches in sequence record
description fields. [default: False]
Outputs:
--o-per-sample-sequences ARTIFACT
SampleData[PairedEndSequencesWithQuality]
The resulting demultiplexed sequences. [required]
--o-error-correction-details ARTIFACT ErrorCorrectionDetails
Detail about the barcode error corrections. [required]
Miscellaneous:
--output-dir PATH Output unspecified results to a directory
--verbose / --quiet Display verbose output to stdout and/or stderr during
execution of this action. Or silence output if
execution is successful (silence is golden).
--example-data PATH Write example data and exit.
--citations Show citations and exit.
--help Show this message and exit.
There were some problems with the command:
(1/2) Invalid value for '--m-barcodes-file': There was an issue with
retrieving column 'sample-id' from the metadata.
(2/2) Invalid value for '--i-seqs': Expected an artifact of at least type
EMPPairedEndSequences. An artifact of type
SampleData[PairedEndSequencesWithQuality] was provided.
(qiime2-metagenome-2024.5) bhagwanrekadwad@MicrobeAI shotgun % qiime demux emp-paired
--m-barcodes-file sample-metadata.tsv
--m-barcodes-column sample-id
--p-rev-comp-barcodes
--i-seqs paired-end-demux-shotgun.qza
--o-per-sample-sequences demux-full.qza
--o-error-correction-details demux-details.qza
Usage: qiime demux emp-paired [OPTIONS]
Demultiplex paired-end sequence data (i.e., map barcode reads to sample ids)
for data generated with the Earth Microbiome Project (EMP) amplicon
sequencing protocol. Details about this protocol can be found at
Protocols and Standards : earthmicrobiome
Inputs:
--i-seqs ARTIFACT EMPPairedEndSequences
The paired-end sequences to be demultiplexed.
[required]
Parameters:
--m-barcodes-file METADATA
--m-barcodes-column COLUMN MetadataColumn[Categorical]
The sample metadata column containing the per-sample
barcodes. [required]
--p-golay-error-correction / --p-no-golay-error-correction
Perform 12nt Golay error correction on the barcode
reads. [default: True]
--p-rev-comp-barcodes / --p-no-rev-comp-barcodes
If provided, the barcode sequence reads will be
reverse complemented prior to demultiplexing.
[default: False]
--p-rev-comp-mapping-barcodes / --p-no-rev-comp-mapping-barcodes
If provided, the barcode sequences in the sample
metadata will be reverse complemented prior to
demultiplexing. [default: False]
--p-ignore-description-mismatch / --p-no-ignore-description-mismatch
If enabled, ignore mismatches in sequence record
description fields. [default: False]
Outputs:
--o-per-sample-sequences ARTIFACT
SampleData[PairedEndSequencesWithQuality]
The resulting demultiplexed sequences. [required]
--o-error-correction-details ARTIFACT ErrorCorrectionDetails
Detail about the barcode error corrections. [required]
Miscellaneous:
--output-dir PATH Output unspecified results to a directory
--verbose / --quiet Display verbose output to stdout and/or stderr during
execution of this action. Or silence output if
execution is successful (silence is golden).
--example-data PATH Write example data and exit.
--citations Show citations and exit.
--help Show this message and exit.
There were some problems with the command:
(1/2) Invalid value for '--m-barcodes-file': There was an issue with
retrieving column 'sample-id' from the metadata.
(2/2) Invalid value for '--i-seqs': Expected an artifact of at least type
EMPPairedEndSequences. An artifact of type
SampleData[PairedEndSequencesWithQuality] was provided.
(qiime2-metagenome-2024.5) bhagwanrekadwad@MicrobeAI shotgun %