vetmohit89 / NanoPsiPy

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no alignment generated #3

Closed eltonjrv closed 2 months ago

eltonjrv commented 2 months ago

Hello,

Thanks for the tool! I'm encountering a problem on the NanoPsiPy_estimation step. It seems that the alignment outputs are not generated, as the "alignment" folder contains only an empty features.csv file, and no "plus_strand" nor "minus_strand" subfolders.

$ NanoPsiPy_estimation -i ../rawData/pus1ko/pus1ko-r1/20240228_1330_MC-116134_FAY49401_30e62174/fastq_pass/ -r ../../refGenome/GQ994935.fasta -o pus1ko_estimation.tsv -s treatment

[M::mm_idx_gen::0.0161.16] collected minimizers [M::mm_idx_gen::0.0261.82] sorted minimizers [M::main::0.0271.79] loaded/built the index for 1 target sequence(s) [M::mm_mapopt_update::0.0291.73] mid_occ = 26 [M::mm_idx_stat] kmer size: 14; skip: 5; is_hpc: 0; #seq: 1 [M::mm_idx_stat::0.031*1.68] distinct minimizers: 45492 (99.22% are singletons); average occurrences: 1.027; average spacing: 2.957; total leng th: 138146 [M::main] Version: 2.18-r1015 [M::main] CMD: minimap2 -ax splice -uf -k14 --secondary=no/../../refGenome/GQ994935 .psu/nanoPsiPy-run/temp/all_pass_fastq.fastq [M::main] Real time: 0.038 sec; CPU: 0.057 sec; Peak RSS: 0.008 GB

Traceback (most recent call last): File "/nobackup/fbsev/bioinformatics-tools/miniconda3/envs/nanoPsiPy/bin/NanoPsiPy_estimation", line 48, in ex_fe() File "/nobackup/fbsev/bioinformatics-tools/miniconda3/envs/nanoPsiPy/lib/python3.11/site-packages/NanoPsiPy/Pseudo_U_estimation.py", line 105, in ex_fe with open(plus_file, "r") as input_file: ^^^^^^^^^^^^^^^^^^^^ FileNotFoundError: [Errno 2] No such file or directory: '.psu/nanoPsiPy-run/alignment/plus_strand/collect_pile_no_intron.txt'

Any light to shed? Thanks

vetmohit89 commented 2 months ago

Have you tested the pipeline with test dataset? I have provided MALAT1 RNA treated with PUS7 enzyme fastq files and MALAT1 sequence.

Try running NanoPsiPy_estimation with this dataset first.

Thank you Mohit

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From: Elton Vasconcelos @.> Sent: Wednesday, May 8, 2024 9:06:15 AM To: vetmohit89/NanoPsiPy @.> Cc: Subscribed @.***> Subject: [vetmohit89/NanoPsiPy] no alignment generated (Issue #3)

Hello,

Thanks for the tool! I'm encountering a problem on the NanoPsiPy_estimation step. It seems that the alignment outputs are not generated, as the "alignment" folder contains only an empty features.csv file, and no "plus_strand" nor "minus_strand" subfolders.

$ NanoPsiPy_estimation -i ../rawData/pus1ko/pus1ko-r1/20240228_1330_MC-116134_FAY49401_30e62174/fastq_pass/ -r ../../refGenome/GQ994935.fasta -o pus1ko_estimation.tsv -s treatment

[M::mm_idx_gen::0.0161.16] collected minimizers [M::mm_idx_gen::0.0261.82] sorted minimizers [M::main::0.0271.79] loaded/built the index for 1 target sequence(s) [M::mm_mapopt_update::0.0291.73] mid_occ = 26 [M::mm_idx_stat] kmer size: 14; skip: 5; is_hpc: 0; #seq: 1 [M::mm_idx_stat::0.031*1.68] distinct minimizers: 45492 (99.22% are singletons); average occurrences: 1.027; average spacing: 2.957; total leng th: 138146 [M::main] Version: 2.18-r1015 [M::main] CMD: minimap2 -ax splice -uf -k14 --secondary=no/../../refGenome/GQ994935 .psu/nanoPsiPy-run/temp/all_pass_fastq.fastq [M::main] Real time: 0.038 sec; CPU: 0.057 sec; Peak RSS: 0.008 GB

Traceback (most recent call last): File "/nobackup/fbsev/bioinformatics-tools/miniconda3/envs/nanoPsiPy/bin/NanoPsiPy_estimation", line 48, in ex_fe() File "/nobackup/fbsev/bioinformatics-tools/miniconda3/envs/nanoPsiPy/lib/python3.11/site-packages/NanoPsiPy/Pseudo_U_estimation.pyhttps://urldefense.com/v3/__http://Pseudo_U_estimation.py__;!!NoSwA-eRAg!BDNGY-FpuwBvxHECYeE6a6Vx8_1wv20UbzemqEjbLgNEHhhjdPboX1yUUAqFcbYtU-Y3tMPh2K_WOK0Ky0t7wu4N$", line 105, in ex_fe with open(plus_file, "r") as input_file: ^^^^^^^^^^^^^^^^^^^^ FileNotFoundError: [Errno 2] No such file or directory: '.psu/nanoPsiPy-run/alignment/plus_strand/collect_pile_no_intron.txt'

Any light to shed? Thanks

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eltonjrv commented 2 months ago

Thanks for your prompt reply. Sorry, my bad! It's working now after I've gunzipped the fastq files. Feel free to close this issue. I'll be running the next steps from your tutorial later today, and will get in further contact if any other issue arises (hopefully not). Thanks again

vetmohit89 commented 2 months ago

You're welcome! Glad to hear it's working now. If you have any further questions, please re-open this issue again.

Thanks