Fast5_to_seq_summary CLI Usage

Activate virtual environment


In [1]:
# Using virtualenvwrapper here but can also be done with Conda 
workon pycoQC


(pycoQC) 

Getting help


In [2]:
Fast5_to_seq_summary -h


usage: Fast5_to_seq_summary [-h] [--version] --fast5_dir FAST5_DIR
                            --seq_summary_fn SEQ_SUMMARY_FN
                            [--max_fast5 MAX_FAST5] [--threads THREADS]
                            [--basecall_id BASECALL_ID]
                            [--fields FIELDS [FIELDS ...]] [--include_path]
                            [--verbose_level VERBOSE_LEVEL]

Fast5_to_seq_summary generate a sequencing summary like file from a directory
containing Fast5 files

optional arguments:
  -h, --help            show this help message and exit
  --version, -v         show program's version number and exit
  --fast5_dir FAST5_DIR, -f FAST5_DIR
                        Directory containing fast5 files. Can contain multiple
                        subdirectories
  --seq_summary_fn SEQ_SUMMARY_FN, -s SEQ_SUMMARY_FN
                        path of the summary sequencing file where to write the
                        data extracted from the fast5 files
  --max_fast5 MAX_FAST5
                        Maximum number of file to try to parse. 0 to
                        deactivate (default: 0)
  --threads THREADS, -t THREADS
                        Total number of threads to use. 1 thread is used for
                        the reader and 1 for the writer. Minimum 3 (default:
                        4)
  --basecall_id BASECALL_ID
                        id of the basecalling group. By default leave to 0,
                        but if you perfome multiple basecalling on the same
                        fast5 files, this can be used to indicate the
                        corresponding group (1, 2 ...) (default: 0)
  --fields FIELDS [FIELDS ...]
                        list of field names corresponding to attributes to try
                        to fetch from the fast5 files (default: ['read_id',
                        'run_id', 'channel', 'start_time',
                        'sequence_length_template', 'mean_qscore_template',
                        'calibration_strand_genome_template',
                        'barcode_arrangement'])
  --include_path        If given, the absolute path to the corresponding file
                        is added in an extra column (default: False)
  --verbose_level VERBOSE_LEVEL
                        Level of verbosity, from 2 (Chatty) to 0 (Nothing)
                        (default: 0)
(pycoQC) 

Usage examples

Basic usage


In [3]:
Fast5_to_seq_summary \
    -f data/ \
    -s results/summary_sequencing.tsv
    
head results/summary_sequencing.tsv


22 reads [00:00, 811.31 reads/s]
Total reads: 22 / Average speed: 543.6 reads/s

(pycoQC) (pycoQC) read_id	run_id	channel	start_time	sequence_length_template	mean_qscore_template	calibration_strand_genome_template
5b7fadd0-c646-4c7b-9800-66ee658a5ca8	40ebe55356ada6c830fa793745ef4c498d896c73	150	37	468	7.608	filtered_out
e6a8e4d0-7b3c-471a-be26-fa7857d12663	40ebe55356ada6c830fa793745ef4c498d896c73	318	15	392	8.304	filtered_out
2c32553e-62c6-4c7a-bf05-249771364f04	40ebe55356ada6c830fa793745ef4c498d896c73	237	11	1151	8.544	filtered_out
f8325de9-a77e-4616-a4a8-69ecf32e1688	40ebe55356ada6c830fa793745ef4c498d896c73	354	16	568	8.206	filtered_out
6af04302-04c8-4d8d-8e87-aa69178b3f24	40ebe55356ada6c830fa793745ef4c498d896c73	36	26	832	8.234	filtered_out
3e81c32a-f2ee-4719-a88d-e0affe93d26f	40ebe55356ada6c830fa793745ef4c498d896c73	348	24	1137	8.124	filtered_out
68804104-71dc-465c-b82d-3a99a4689701	40ebe55356ada6c830fa793745ef4c498d896c73	38	20	1010	8.325	filtered_out
37dfa1d5-5d84-486c-bf47-9eb6438f5645	40ebe55356ada6c830fa793745ef4c498d896c73	410	30	555	8.219	filtered_out
3784283c-47cc-48ac-8d7b-7efd32123b56	40ebe55356ada6c830fa793745ef4c498d896c73	243	20	893	8.54	filtered_out
(pycoQC) 

Multi-threading support


In [4]:
Fast5_to_seq_summary \
    -f data/ \
    -s results/summary_sequencing.tsv \
    -t 6
    
head results/summary_sequencing.tsv


22 reads [00:00, 1129.77 reads/s]
Total reads: 22 / Average speed: 650.82 reads/s

(pycoQC) (pycoQC) read_id	run_id	channel	start_time	sequence_length_template	mean_qscore_template	calibration_strand_genome_template
5b7fadd0-c646-4c7b-9800-66ee658a5ca8	40ebe55356ada6c830fa793745ef4c498d896c73	150	37	468	7.608	filtered_out
e6a8e4d0-7b3c-471a-be26-fa7857d12663	40ebe55356ada6c830fa793745ef4c498d896c73	318	15	392	8.304	filtered_out
f8325de9-a77e-4616-a4a8-69ecf32e1688	40ebe55356ada6c830fa793745ef4c498d896c73	354	16	568	8.206	filtered_out
2c32553e-62c6-4c7a-bf05-249771364f04	40ebe55356ada6c830fa793745ef4c498d896c73	237	11	1151	8.544	filtered_out
6af04302-04c8-4d8d-8e87-aa69178b3f24	40ebe55356ada6c830fa793745ef4c498d896c73	36	26	832	8.234	filtered_out
68804104-71dc-465c-b82d-3a99a4689701	40ebe55356ada6c830fa793745ef4c498d896c73	38	20	1010	8.325	filtered_out
3e81c32a-f2ee-4719-a88d-e0affe93d26f	40ebe55356ada6c830fa793745ef4c498d896c73	348	24	1137	8.124	filtered_out
37dfa1d5-5d84-486c-bf47-9eb6438f5645	40ebe55356ada6c830fa793745ef4c498d896c73	410	30	555	8.219	filtered_out
3784283c-47cc-48ac-8d7b-7efd32123b56	40ebe55356ada6c830fa793745ef4c498d896c73	243	20	893	8.54	filtered_out
(pycoQC) 

Customize fields of the summary file


In [5]:
Fast5_to_seq_summary \
    -f data/ \
    -s results/custom_summary_sequencing.tsv \
    --verbose_level 1 \
    --fields mean_qscore_template called_events duration strand_score
    
head results/custom_summary_sequencing.tsv


Check input data and options
Start processing fast5 files
22 reads [00:00, 821.28 reads/s]
Overall counts 	valid files: 22

fields found 	mean_qscore_template: 22
	called_events: 22
	duration: 22
	strand_score: 22

fields not found 
Total reads: 22 / Average speed: 464.09 reads/s

(pycoQC) (pycoQC) mean_qscore_template	called_events	duration	strand_score
8.304	1547	23218	-0.0008
8.325	3846	57697	-0.0004
7.608	1615	24233	-0.0007
8.219	2080	31208	-0.0011
7.805	1438	21583	-0.0006
8.544	3740	56107	-0.0003
8.23	2778	51387	-0.0007
8.741	3117	46767	-0.0006
8.987	4235	63526	-0.0003
(pycoQC) 

Add file path


In [6]:
Fast5_to_seq_summary \
    -f data/ \
    -s results/fn_summary_sequencing.tsv \
    --verbose_level 1 \
    --include_path
    
head results/fn_summary_sequencing.tsv


Check input data and options
Start processing fast5 files
22 reads [00:00, 493.99 reads/s]
Overall counts 	valid files: 22

fields found 	read_id: 22
	run_id: 22
	channel: 22
	start_time: 22
	sequence_length_template: 22
	mean_qscore_template: 22
	calibration_strand_genome_template: 22

fields not found 	barcode_arrangement: 22

Total reads: 22 / Average speed: 364.14 reads/s

(pycoQC) (pycoQC) read_id	run_id	channel	start_time	sequence_length_template	mean_qscore_template	calibration_strand_genome_template	path
5b7fadd0-c646-4c7b-9800-66ee658a5ca8	40ebe55356ada6c830fa793745ef4c498d896c73	150	37	468	7.608	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_150_strand.fast5
2c32553e-62c6-4c7a-bf05-249771364f04	40ebe55356ada6c830fa793745ef4c498d896c73	237	11	1151	8.544	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_237_strand.fast5
e6a8e4d0-7b3c-471a-be26-fa7857d12663	40ebe55356ada6c830fa793745ef4c498d896c73	318	15	392	8.304	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_318_strand.fast5
f8325de9-a77e-4616-a4a8-69ecf32e1688	40ebe55356ada6c830fa793745ef4c498d896c73	354	16	568	8.206	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_354_strand.fast5
3e81c32a-f2ee-4719-a88d-e0affe93d26f	40ebe55356ada6c830fa793745ef4c498d896c73	348	24	1137	8.124	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_348_strand.fast5
37dfa1d5-5d84-486c-bf47-9eb6438f5645	40ebe55356ada6c830fa793745ef4c498d896c73	410	30	555	8.219	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_410_strand.fast5
9a1c5296-2ab1-4abd-8d50-e059754cf332	40ebe55356ada6c830fa793745ef4c498d896c73	319	33	1235	8.119	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_319_strand.fast5
6af04302-04c8-4d8d-8e87-aa69178b3f24	40ebe55356ada6c830fa793745ef4c498d896c73	36	26	832	8.234	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_36_strand.fast5
0ef36fa0-7c3f-47f5-8410-027f50724701	40ebe55356ada6c830fa793745ef4c498d896c73	108	26	915	9.407	filtered_out	/home/aleg/Programming/Packages/pycoQC/docs/Fast5_to_seq_summary/data/20180625_FAH77625_MN23126_sequencing_run_S1_57529_read_10_ch_108_strand.fast5
(pycoQC)