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rampart with dorado's .fastq #117

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doloacu opened this issue Sep 18, 2024 · 1 comment
Open

rampart with dorado's .fastq #117

doloacu opened this issue Sep 18, 2024 · 1 comment

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@doloacu
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doloacu commented Sep 18, 2024

Hello, for first time I am having some problems since I started to use rampart after generate my fastq with dorado. I have this kind of problems:
(artic-rampart) minion@minion-H470M-DS3H:~/Documents/20240916/rampart-rsv$ rampart --protocol . --basecalledPath /home/minion/Documents/20240916/demux-dorado-fast
Creating path /home/minion/Documents/20240916/rampart-rsv/annotations/


RAMPART daemon running
socket open on port 3001 for all data communication


Serving built bundle at http://localhost:3000

Found 0 annotated CSV files in .../rampart-rsv/annotations/. FASTQs with the same filename as these will be ignored.
Scanning folder /home/minion/Documents/20240916/demux-dorado-fast/ for FASTQs
(basecalled files which exist here (or are created here by MinKNOW) will be annotated and loaded)

Initial scan for FASTQs complete. Found 88 files, 88 of which are unannotated (unprocesed).
Watching for new files
[warning] pipeline (Annotate reads) finished with exit code 1. Error messages:
[warning] Building DAG of jobs...

[warning] Relative file path './references.fasta' starts with './'. This is redundant and strongly discouraged. It can also lead to inconsistent results of the file-matching approach used by Snakemake. You can simply omit the './' for relative file paths.

[warning] Relative file path './references.fasta' starts with './'. This is redundant and strongly discouraged. It can also lead to inconsistent results of the file-matching approach used by Snakemake. You can simply omit the './' for relative file paths.

[warning] Using shell: /usr/bin/bash
Provided cores: 2

[warning] Rules claiming more threads will be scaled down.

[warning] Job counts:
count jobs
1 all
1 minimap2
1 parse_mapping
3

[warning]
[Wed Sep 18 13:39:03 2024]

[warning] rule minimap2:
input: /home/minion/Documents/20240916/demux-dorado-fast/SQK-RBK110-96_barcode01.fastq, ./references.fasta
output: /home/minion/Documents/20240916/rampart-rsv/annotations/temp/SQK-RBK110-96_barcode01.paf
jobid: 2
wildcards: filename_stem=SQK-RBK110-96_barcode01
threads: 2

[warning]

[warning] [M::mm_idx_gen::0.001*1.45] collected minimizers

[warning] [M::mm_idx_gen::0.0021.67] sorted minimizers
[M::main::0.002
1.66] loaded/built the index for 1 target sequence(s)

[warning] [M::mm_mapopt_update::0.002*1.61] mid_occ = 3
[M::mm_idx_stat] kmer size: 15; skip: 10; is_hpc: 0; #seq: 1

[warning] [M::mm_idx_stat::0.002*1.57] distinct minimizers: 5543 (97.44% are singletons); average occurrences: 1.026; average spacing: 5.419

[warning] [M::worker_pipeline::3.230*1.92] mapped 76470 sequences

[warning] [M::main] Version: 2.17-r941
[M::main] CMD: minimap2 -t 2 -x map-ont --secondary=no --paf-no-hit --cs ./references.fasta /home/minion/Documents/20240916/demux-dorado-fast/SQK-RBK110-96_barcode01.fastq
[M::main] Real time: 3.230 sec; CPU: 6.188 sec; Peak RSS: 0.091 GB

[warning] [Wed Sep 18 13:39:06 2024]

[warning] Finished job 2.

[warning] 1 of 3 steps (33%) done

[warning]
[Wed Sep 18 13:39:06 2024]

[warning] rule parse_mapping:
input: /home/minion/Documents/20240916/demux-dorado-fast/SQK-RBK110-96_barcode01.fastq, /home/minion/Documents/20240916/rampart-rsv/annotations/temp/SQK-RBK110-96_barcode01.paf, ./references.fasta
output: /home/minion/Documents/20240916/rampart-rsv/annotations/SQK-RBK110-96_barcode01.csv
jobid: 1
wildcards: filename_stem=SQK-RBK110-96_barcode01

[warning]

[warning] Traceback (most recent call last):
File "/home/minion/rampart/default_protocol/pipelines/demux_map/rules/parse_paf.py", line 83, in get_header_dict
barcode = header["barcode"]
KeyError: 'barcode'

During handling of the above exception, another exception occurred:

Traceback (most recent call last):
File "/home/minion/rampart/default_protocol/pipelines/demux_map/rules/parse_paf.py", line 288, in
header_dict = get_header_dict(args.reads)
File "/home/minion/rampart/default_protocol/pipelines/demux_map/rules/parse_paf.py", line 87, in get_header_dict
start_time = header["start_time"]
KeyError: 'start_time'

[warning] [Wed Sep 18 13:39:06 2024]

[warning] Error in rule parse_mapping:
jobid: 1

[warning] output: /home/minion/Documents/20240916/rampart-rsv/annotations/SQK-RBK110-96_barcode01.csv

[warning] shell:

    python /home/minion/rampart/default_protocol/pipelines/demux_map/rules/parse_paf.py         --paf_file /home/minion/Documents/20240916/rampart-rsv/annotations/temp/SQK-RBK110-96_barcode01.paf         --report /home/minion/Documents/20240916/rampart-rsv/annotations/SQK-RBK110-96_barcode01.csv         --annotated_reads /home/minion/Documents/20240916/demux-dorado-fast/SQK-RBK110-96_barcode01.fastq         --reference_file ./references.fasta                  --reference_options ""
    
    (one of the commands exited with non-zero exit code; note that snakemake uses bash strict mode!)
@rmcolq
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rmcolq commented Sep 19, 2024

Does Dorado still include the start_time in the header of reads? Or more generally, what does the header line for a read in the output fastq file look like by comparison with e.g. guppy?

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