Have you encountered this error while Denoising using DADA2

Technical Support
1

demux.qzv (313.3 KB)

qiime dada2 denoise-paired --i-demultiplexed-seqs imported-paired-end-seqs.qza --p-trim-left-f 0 --p-trunc-len-f 270 --p-trim-left-r 0 --p-trunc-len-r 240 --o-representative-sequences rep-seqs.qza --o-table table.qza --o-denoising-stats denoising-stats.qza --p-n-threads 0 --verbose
Running external command line application(s). This may print messages to stdout and/or stderr.
The command(s) being run are below. These commands cannot be manually re-run as they will depend on temporary files that no longer exist.

Command: run_dada.R --input_directory /tmp/tmpuocii1eb/forward --input_directory_reverse /tmp/tmpuocii1eb/reverse --output_path /tmp/tmpuocii1eb/output.tsv.biom --output_track /tmp/tmpuocii1eb/track.tsv --filtered_directory /tmp/tmpuocii1eb/filt_f --filtered_directory_reverse /tmp/tmpuocii1eb/filt_r --truncation_length 270 --truncation_length_reverse 240 --trim_left 0 --trim_left_reverse 0 --max_expected_errors 2.0 --max_expected_errors_reverse 2.0 --truncation_quality_score 2 --min_overlap 12 --pooling_method independent --chimera_method consensus --min_parental_fold 1.0 --allow_one_off False --num_threads 0 --learn_min_reads 1000000

Warning message:
package ‘optparse’ was built under R version 4.2.3
R version 4.2.2 (2022-10-31)
Loading required package: Rcpp
DADA2: 1.26.0 / Rcpp: 1.0.12 / RcppParallel: 5.1.6
2) Filtering Error in names(answer) <- names1 :
'names' attribute [13] must be the same length as the vector [9]
4: mcmapply(fastqPairedFilter, mapply(c, fwd, rev, SIMPLIFY = FALSE),
mapply(c, filt, filt.rev, SIMPLIFY = FALSE), MoreArgs = list(truncQ = truncQ,
truncLen = truncLen, trimLeft = trimLeft, trimRight = trimRight,
maxLen = maxLen, minLen = minLen, maxN = maxN, minQ = minQ,
maxEE = maxEE, rm.phix = rm.phix, rm.lowcomplex = rm.lowcomplex,
orient.fwd = orient.fwd, matchIDs = matchIDs, id.sep = id.sep,
id.field = id.field, n = n, OMP = OMP, qualityType = qualityType,
compress = compress, verbose = verbose), mc.cores = ncores,
mc.silent = TRUE)
3: filterAndTrim(unfilts, filts, unfiltsR, filtsR, truncLen = c(truncLen,
truncLenR), trimLeft = c(trimLeft, trimLeftR), maxEE = c(maxEE,
maxEER), truncQ = truncQ, rm.phix = TRUE, multithread = multithread)
2: withCallingHandlers(expr, warning = function(w) if (inherits(w,
classes)) tryInvokeRestart("muffleWarning"))
1: suppressWarnings(filterAndTrim(unfilts, filts, unfiltsR, filtsR,
truncLen = c(truncLen, truncLenR), trimLeft = c(trimLeft,
trimLeftR), maxEE = c(maxEE, maxEER), truncQ = truncQ,
rm.phix = TRUE, multithread = multithread))
Traceback (most recent call last):
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/site-packages/q2_dada2/_denoise.py", line 350, in denoise_paired
run_commands([cmd])
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/site-packages/q2_dada2/_denoise.py", line 37, in run_commands
subprocess.run(cmd, check=True)
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/subprocess.py", line 516, in run
raise CalledProcessError(retcode, process.args,
subprocess.CalledProcessError: Command '['run_dada.R', '--input_directory', '/tmp/tmpuocii1eb/forward', '--input_directory_reverse', '/tmp/tmpuocii1eb/reverse', '--output_path', '/tmp/tmpuocii1eb/output.tsv.biom', '--output_track', '/tmp/tmpuocii1eb/track.tsv', '--filtered_directory', '/tmp/tmpuocii1eb/filt_f', '--filtered_directory_reverse', '/tmp/tmpuocii1eb/filt_r', '--truncation_length', '270', '--truncation_length_reverse', '240', '--trim_left', '0', '--trim_left_reverse', '0', '--max_expected_errors', '2.0', '--max_expected_errors_reverse', '2.0', '--truncation_quality_score', '2', '--min_overlap', '12', '--pooling_method', 'independent', '--chimera_method', 'consensus', '--min_parental_fold', '1.0', '--allow_one_off', 'False', '--num_threads', '0', '--learn_min_reads', '1000000']' returned non-zero exit status 1.

During handling of the above exception, another exception occurred:

Traceback (most recent call last):
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/site-packages/q2cli/commands.py", line 520, in call
results = self._execute_action(
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/site-packages/q2cli/commands.py", line 581, in _execute_action
results = action(**arguments)
File "", line 2, in denoise_paired
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/site-packages/qiime2/sdk/action.py", line 342, in bound_callable
outputs = self.callable_executor(
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/site-packages/qiime2/sdk/action.py", line 566, in callable_executor
output_views = self._callable(**view_args)
File "/home/nemalab307/anaconda3/envs/qiime2-amplicon-2024.2/lib/python3.8/site-packages/q2_dada2/_denoise.py", line 363, in denoise_paired
raise Exception("An error was encountered while running DADA2"
Exception: An error was encountered while running DADA2 in R (return code 1), please inspect stdout and stderr to learn more.

Plugin error from dada2:

An error was encountered while running DADA2 in R (return code 1), please inspect stdout and stderr to learn more.

See above for debug info.

3

Hello @rpaudel. The key part of the traceback is

which isn't very informative. Fortunately, we have encountered this error a number of times, and have determined that it is effectively an "out of memory" error even though it doesn't look like one. Try running the action with --p-n-threads 1. It will take longer but it will minimize memory usage.

It is possible that you can get away with using more than 1 thread if you want to play with the number and find the sweet spot, but if 1 doesn't work then every other number will also run out of memory. 0 is using as many threads as you have CPU threads.

1 Like
5

Thank you! It worked.

1 Like