Dada2 error message

[drmusk (Muhammad Umar)]

Dear All
I have 38 paired end fastq files while I have uploaded to paired-end-demux.qza through manifest file. Now I wanted to apply Dada2 but I am getting error.

I am applying this command in qiime2019.4

qiime dada2 denoise-paired
–i-demultiplexed-seqs paired-end-demux.qza
–p-trim-left-f 10
–p-trim-left-r 10
–p-trunc-len-f 300
–p-trunc-len-r 300
–o-table table.qza
–o-representative-sequences rep-seqs.qza
–o-denoising-stats denoising-stats.qza
–p-n-threads 0

but I am getting below error message.

There was a problem with the command:
(1/1) Invalid value for “–i-demultiplexed-seqs”: ‘paired-end-demux.qza’ is
not a QIIME 2 Artifact (.qza)
Traceback (most recent call last):
File “/home/qiime2/miniconda/envs/qiime2-2019.4/lib/python3.6/weakref.py”, line 624, in _exitfunc
f()
File “/home/qiime2/miniconda/envs/qiime2-2019.4/lib/python3.6/weakref.py”, line 548, in call
return info.func(info.args, *(info.kwargs or {}))
File “/home/qiime2/miniconda/envs/qiime2-2019.4/lib/python3.6/site-packages/qiime2/core/path.py”, line 99, in _destruct
shutil.rmtree(path)
File “/home/qiime2/miniconda/envs/qiime2-2019.4/lib/python3.6/shutil.py”, line 480, in rmtree
_rmtree_safe_fd(fd, path, onerror)
File “/home/qiime2/miniconda/envs/qiime2-2019.4/lib/python3.6/shutil.py”, line 418, in _rmtree_safe_fd
_rmtree_safe_fd(dirfd, fullname, onerror)
File “/home/qiime2/miniconda/envs/qiime2-2019.4/lib/python3.6/shutil.py”, line 438, in _rmtree_safe_fd
onerror(os.unlink, fullname, sys.exc_info())
File “/home/qiime2/miniconda/envs/qiime2-2019.4/lib/python3.6/shutil.py”, line 436, in _rmtree_safe_fd
os.unlink(name, dir_fd=topfd)
OSError: [Errno 30] Read-only file system: ‘metadata.yaml’

[Mehrbod_Estaki]

Hi @drmusk,
Can you double check to make sure there is in fact a paired-end-demux.qza in the directory you are working in, double check the exact name too. This is the most common source of this error, an incorrect file path.

[drmusk (Muhammad Umar)]

Yes Mehrbod. You are right. Actually I made paired-end-demux.qza in one computer and moved it to other computer for further processing and that might resulted in a change in manifest file oriented paired-end-demux.qza sequences.
When I again started working in first computer it went well.