Program options for guppy_basecaller: : Guppy Basecalling Software, (C) Oxford Nanopore Technologies, Limited. Version 4.0.15+5694074, client-server API version 2.1.0 Usage: With config file:" guppy_basecaller -i -s -c [options] With flowcell and kit name: guppy_basecaller -i -s --flowcell --kit List supported flowcells and kits: guppy_basecaller --print_workflows Use GPU for basecalling: guppy_basecaller -i -s -c --device [options] Use server for basecalling: guppy_basecaller -i -s -c --port [options] Command line parameters: --trim_threshold arg Threshold above which data will be trimmed (in standard deviations of current level distribution). --trim_min_events arg Adapter trimmer minimum stride intervals after stall that must be seen. --max_search_len arg Maximum number of samples to search through for the stall --override_scaling Manually provide scaling parameters rather than estimating them from each read. --scaling_med arg Median current value to use for manual scaling. --scaling_mad arg Median absolute deviation to use for manual scaling. --trim_strategy arg Trimming strategy to apply: 'dna' or 'rna' (or 'none' to disable trimming) --dmean_win_size arg Window size for coarse stall event detection --dmean_threshold arg Threshold for coarse stall event detection --jump_threshold arg Threshold level for rna stall detection --pt_scaling Enable polyT/adapter max detection for read scaling. --pt_median_offset arg Set polyT median offset for setting read scaling median (default 2.5) --adapter_pt_range_scale arg Set polyT/adapter range scale for setting read scaling median absolute deviation (default 5.2) --pt_required_adapter_drop arg Set minimum required current drop from adapter max to polyT detection. (default 30.0) --pt_minimum_read_start_index arg Set minimum index for read start sample required to attempt polyT scaling. (default 30) --as_model_file arg Path to JSON model file for adapter scaling. --as_gpu_runners_per_device arg Number of runners per GPU device for adapter scaling. --as_cpu_threads_per_scaler arg Number of CPU worker threads per adapter scaler --as_reads_per_runner arg Maximum reads per runner for adapter scaling. --as_num_scalers arg Number of parallel scalers for adapter scaling. -m [ --model_file ] arg Path to JSON model file. -k [ --kernel_path ] arg Path to GPU kernel files location (only needed if builtin_scripts is false). -x [ --device ] arg Specify basecalling device: 'auto', or 'cuda:'. --builtin_scripts arg Whether to use GPU kernels that were included at compile-time. --chunk_size arg Stride intervals per chunk. --chunks_per_runner arg Maximum chunks per runner. --chunks_per_caller arg Soft limit on number of chunks in each caller's queue. New reads will not be queued while this is exceeded. --high_priority_threshold arg Number of high priority chunks to process for each medium priority chunk. --medium_priority_threshold arg Number of medium priority chunks to process for each low priority chunk. --overlap arg Overlap between chunks (in stride intervals). --gpu_runners_per_device arg Number of runners per GPU device. --cpu_threads_per_caller arg Number of CPU worker threads per basecaller. --num_callers arg Number of parallel basecallers to create. --post_out Return full posterior matrix in output fast5 file and/or called read message from server. --stay_penalty arg Scaling factor to apply to stay probability calculation during transducer decode. --qscore_offset arg Qscore calibration offset. --qscore_scale arg Qscore calibration scale factor. --temp_weight arg Temperature adjustment for weight matrix in softmax layer of RNN. --temp_bias arg Temperature adjustment for bias vector in softmax layer of RNN. --qscore_filtering Enable filtering of reads into PASS/FAIL folders based on min qscore. --min_qscore arg Minimum acceptable qscore for a read to be filtered into the PASS folder --reverse_sequence arg Reverse the called sequence (for RNA sequencing). --u_substitution arg Substitute 'U' for 'T' in the called sequence (for RNA sequencing). --log_speed_frequency arg How often to print out basecalling speed. --barcode_kits arg Space separated list of barcoding kit(s) or expansion kit(s) to detect against. Must be in double quotes. --trim_barcodes Trim the barcodes from the output sequences in the FastQ files. --num_extra_bases_trim arg How vigorous to be in trimming the barcode. Default is 0 i.e. the length of the detected barcode. A positive integer means extra bases will be trimmed, a negative number is how many fewer bases (less vigorous) will be trimmed. --arrangements_files arg Files containing arrangements. --score_matrix_filename arg File containing mismatch score matrix. --start_gap1 arg Gap penalty for aligning before the reference. --end_gap1 arg Gap penalty for aligning after the reference. --open_gap1 arg Penalty for opening a new gap in the reference. --extend_gap1 arg Penalty for extending a gap in the reference. --start_gap2 arg Gap penalty for aligning before the query. --end_gap2 arg Gap penalty for aligning after the query. --open_gap2 arg Penalty for opening a new gap in the query. --extend_gap2 arg Penalty for extending a gap in the query. --min_score arg Minimum score to consider a valid alignment. --min_score_rear_override arg Minimum score to consider a valid alignment for the rear barcode only (and min_score will then be used for the front only when this is set). --front_window_size arg Window size for the beginning barcode. --rear_window_size arg Window size for the ending barcode. --require_barcodes_both_ends Reads will only be classified if there is a barcode above the min_score at both ends of the read. --allow_inferior_barcodes Reads will still be classified even if both the barcodes at the front and rear (if applicable) were not the best scoring barcodes above the min_score. --detect_mid_strand_barcodes Search for barcodes through the entire length of the read. --min_score_mid_barcodes arg Minimum score for a barcode to be detected in the middle of a read. --num_barcoding_buffers arg Number of GPU memory buffers to allocate to perform barcoding into. Controls level of parallelism on GPU for barcoding. --num_barcode_threads arg Number of worker threads to use for barcoding. --calib_detect Enable calibration strand detection and filtering. --calib_reference arg Reference FASTA file containing calibration strand. --calib_min_sequence_length arg Minimum sequence length for reads to be considered candidate calibration strands. --calib_max_sequence_length arg Maximum sequence length for reads to be considered candidate calibration strands. --calib_min_coverage arg Minimum reference coverage to pass calibration strand detection. --print_workflows Output available workflows. --flowcell arg Flowcell to find a configuration for --kit arg Kit to find a configuration for -a [ --align_ref ] arg Path to alignment reference. --bed_file arg Path to .bed file containing areas of interest in reference genome. --num_alignment_threads arg Number of worker threads to use for alignment. -z [ --quiet ] Quiet mode. Nothing will be output to STDOUT if this option is set. --trace_categories_logs arg Enable trace logs - list of strings with the desired names. --verbose_logs Enable verbose logs. --trace_domains_log arg List of trace domains to include in verbose logging (if enabled), '*' for all. --trace_domains_config arg Configuration file containing list of trace domains to include in verbose logging (if enabled), this will override --trace_domain_logs --disable_pings Disable the transmission of telemetry pings. --ping_url arg URL to send pings to --ping_segment_duration arg Duration in minutes of each ping segment. --progress_stats_frequency arg Frequency in seconds in which to report progress statistics, if supplied will replace the default progress display. -q [ --records_per_fastq ] arg Maximum number of records per fastq file, 0 means use a single file (per worker, per run id). --read_batch_size arg Maximum batch size, in reads, for grouping input files. --compress_fastq Compress fastq output files with gzip. -i [ --input_path ] arg Path to input fast5 files. --input_file_list arg Optional file containing list of input fast5 files to process from the input_path. -s [ --save_path ] arg Path to save fastq files. -l [ --read_id_list ] arg File containing list of read ids to filter to -r [ --recursive ] Search for input files recursively. --fast5_out Choice of whether to do fast5 output. --resume Resume a previous basecall run using the same output folder. --max_block_size arg Maximum block size (in events) of basecall messages to server. -p [ --port ] arg Port for basecalling service. --barcoding_config_file arg Configuration file to use for barcoding. --disable_events Disable the transmission of event tables when receiving reads back from the basecall server. --client_id arg Optional unique identifier (non-negative integer) for this instance of the Guppy Client Basecaller, if supplied will form part of the output filenames. --nested_output_folder If flagged output fastq files will be written to a nested folder structure, based on: protocol_group/sample/protocol/qscore_p ass_fail/barcode_arrangement/ --server_file_load_timeout arg Timeout in seconds to wait for the server to load a requested data file (e.g. a basecalling model or alignment index). default is 30s --max_queued_reads arg Maximum number of reads to be submitted for processing at any one time. -h [ --help ] produce help message -v [ --version ] print version number -c [ --config ] arg Config file to use -d [ --data_path ] arg Path to use for loading any data files the application requires. ----------------------------------------------