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a-h-b avatar a-h-b commented on August 23, 2024

Hi -
thanks for getting in touch. I think you've indeed missed something when installing or just got the command for the initialisation wrong. If I got you right, you tried
/home/anil/dadasnake/submit_scripts/dadasnake_withReport.sh -i /home/anil/dadasnake/config/config.init.yaml
when indeed, you should call the dadasnake script:
/home/anil/dadasnake/dadasnake -i /home/anil/dadasnake/config/config.init.yaml `
This script isn't there when you clone the pipeline from github. You copy it from either of the auxiliary_files/dadasnake_* files, as described in step 3.
I hope this helps. Please let me know, if there are more questions.
-A

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

Thank you for your quick response.
I realized that i skipped some of the steps of installation as i was unable to understand some of the terms as i am a wet lab student and very recently i started learning all this dry lab work.

as suggested i ran the above command and got a new issue
/home/anil/dadasnake/dadasnake -i /home/anil/dadasnake/config/config.init.yaml
Initializing conda environments.
module: command not found
Traceback (most recent call last):
File "/home/anil/dadasnake/conda/snakemake_env/bin/snakemake", line 10, in
sys.exit(main())
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/snakemake/init.py", line 2592, in main
success = snakemake(
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/snakemake/init.py", line 504, in snakemake
overwrite_config.update(load_configfile(f))
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/snakemake/io.py", line 1596, in load_configfile
raise WorkflowError(
snakemake.exceptions.WorkflowError: Config file must be given as JSON or YAML with keys at top level.

Please help

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a-h-b avatar a-h-b commented on August 23, 2024

Hi -
glad we could straighten that one out.
yes, for your new error, you need to change the VARIABLE_CONFIG file, particularly in the line that currently reads:
LOADING_MODULES module load Anaconda3/5.3.0
it should just be
LOADING_MODULES
because it seems your system does not rely on modules.
Let me know, if there are other errors.
Best wishes -
A

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

sorry for the constant trouble
i edited the VARIABLE_CONFIG file as suggested but still got this error
/home/anil/dadasnake/dadasnake -i /home/anil/dadasnake/config/config.init.yaml
Initializing conda environments.
Traceback (most recent call last):
File "/home/anil/dadasnake/conda/snakemake_env/bin/snakemake", line 10, in
sys.exit(main())
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/snakemake/init.py", line 2592, in main
success = snakemake(
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/snakemake/init.py", line 504, in snakemake
overwrite_config.update(load_configfile(f))
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/snakemake/io.py", line 1596, in load_configfile
raise WorkflowError(
snakemake.exceptions.WorkflowError: Config file must be given as JSON or YAML with keys at top level

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a-h-b avatar a-h-b commented on August 23, 2024

Three tests, please:
a) how did you install snakemake?
b) can you please post the output of
/home/anil/dadasnake/dadasnake -h
c) I guess you did not change anything in /home/anil/dadasnake/config/config.init.yaml ?
Thanks -
A

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

a) i installed snakemake using step 4 of the guide as
conda install -c conda-forge mamba
mamba create --prefix /home/anil/dadasnake/conda/snakemake_env
conda activate /home/anil/dadasnake/conda/snakemake_env
mamba install -c conda-forge -c bioconda snakemake
conda deactivate

b)/home/anil/dadasnake/dadasnake -h
Usage: /home/anil/dadasnake/dadasnake [-u|d|c|l|i] [-r] [-t number] [-n name] /absolute_path/to/config_file
-n , only works with -c
-r is set, a report is generated (it's recommended to run -c or -l with -r)
-d if set, a dryrun is performed
-c if set, the whole thing is submitted to the cluster
-i if set, only the conda environments will be installed, if they don't exist
-u if set, the working directory will be unlocked (only necessary for crash/kill recovery)
-l if set, the main snakemake thread and indivdual rules are run in the current terminal session
-t <max_threads> maximum number of cpus to use for all rules at a time. Defaults to 50 for -c, and to 1 for -l. No effect on -r, -d or -u only.
c) yes i did change some in the config.init.yaml file
outputdir dadasnake_initialized
taxonomy
mothur
do true
blast
do true
hand_off
biom true
phyloseq true

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a-h-b avatar a-h-b commented on August 23, 2024

do you mean to say you deleted the ":"s ?

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a-h-b avatar a-h-b commented on August 23, 2024

and the spaces at the start of the lines?

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

No i only removed : and ":" not the spaces at the start of the line

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a-h-b avatar a-h-b commented on August 23, 2024

okay, that explains the error. Please don't remove the : - they're part of the yaml format.
-A

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

ok as suggested i added the : in the file
still got this
/home/anil/dadasnake/dadasnake -i /home/anil/dadasnake/config/config.init.yaml
Initializing conda environments.

Final resource settings:
maxCores: 1
Sample table was not found. Please enter the absolute path and file name in the config file.
FileNotFoundError in line 113 of /home/anil/dadasnake/workflow/rules/get_config.smk:
[Errno 2] No such file or directory: '/home/anil/testdata/samples.small.tsv'
File "/home/anil/dadasnake/Snakefile", line 8, in
File "/home/anil/dadasnake/workflow/rules/get_config.smk", line 113, in
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/pandas/io/parsers.py", line 689, in read_table
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/pandas/io/parsers.py", line 462, in _read
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/pandas/io/parsers.py", line 819, in init
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/pandas/io/parsers.py", line 1050, in _make_engine
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/pandas/io/parsers.py", line 1867, in init
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/pandas/io/parsers.py", line 1362, in _open_handles
File "/home/anil/dadasnake/conda/snakemake_env/lib/python3.9/site-packages/pandas/io/common.py", line 642, in get_handle

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

also i am not running dadasnake in the cluster but on a ubuntu desktop so which dadasnake file should i copy in the step 3 from auxiliary_files.
i copied the one with the process running snakemake to the cluster

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a-h-b avatar a-h-b commented on August 23, 2024

Regarding your second question: it shouldn't make a difference in that case. For the proper runs, you'd be using the -l setting then.
I am still trying to figure out why the workflow is looking in the wrong directory. You didn't change the dadasnake file, did you? nor the config/config.default.yaml?
-A

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

i guess you are asking me about this file /home/anil/dadasnake/dadasnake if that's the case i didn"t change anything in this file nor in the config/config.default.yaml file

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a-h-b avatar a-h-b commented on August 23, 2024

ok

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a-h-b avatar a-h-b commented on August 23, 2024

oh, I am sorry, I did not describe this well in the README. Can you please do
cd /home/anil/dadasnake/
and then run the initialisation from there?
i.e.
./dadasnake -i config/config.init.yaml

thanks -
A

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

ya now the command is running untill it runs i have some questions

  1. can i process my 16s nanopore reads using the pipeline
  2. in the database downloading steps you suggested to download database for mothur, DADA2 implementation database from McLaren, database for DECIPHER, database for fungal traits
    do i need to download all these databases....?

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a-h-b avatar a-h-b commented on August 23, 2024
  1. ha, well, we've set up a workflow that should be able to work on ChopSeq-treated data from Nanopore sequencing - this is from a consensus sequencing method (https://academic.oup.com/gigascience/article/7/12/giy140/5202451). It shouldn't work native Nanopore reads, though: benjjneb/dada2#718
  2. no, you just need the database that suits your needs (e.g. just DECIPHER, if your dataset is quite small; or just the SILVA data base for mothur) assuming you want to only run 16S data

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

ok, so u meant that to analyse the nanopore reads i have to firstly process my reads using chopseq and then run the dadasnake right...?

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a-h-b avatar a-h-b commented on August 23, 2024

if you've sequenced them with the method described in that paper or the INCseq method. If you've just sequenced amplicons without circular consensus, please use the Nanopore (what's in my pot?) workflow for processing.

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

okay
The command ran smoothly
Thank you for your support

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

if you've sequenced them with the method described in that paper or the INCseq method. If you've just sequenced amplicons without circular consensus, please use the Nanopore (what's in my pot?) workflow for processing.

okay thank you for the information

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a-h-b avatar a-h-b commented on August 23, 2024

you're welcome. Good luck with your analysis.
-A

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anilchauhanhp9 avatar anilchauhanhp9 commented on August 23, 2024

Thanks

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