Quality control of reads¶

Adapter Trimming¶

FASTA file paths for adapter sequences to be trimmed from the sequence ends.

We provide the adapter reference FASTA included in bbmap for various

preprocess_adapters: /database_dir/adapters.fa

Trim regions with an average quality below this threshold. Higher is more stringent.

preprocess_minimum_base_quality: 10

Allow shorter kmer matches down to mink at the read ends. 0 disables.

preprocess_adapter_min_k: 8

Maximum number of substitutions between the target adapter kmer and the query sequence kmer. Lower is more stringent.

preprocess_allowable_kmer_mismatches: 1

Kmer length used for finding contaminants. Contaminant matches shorter than this length will not be found.

preprocess_reference_kmer_match_length: 27

This is applied after quality and adapter trimming have been applied to the sequence.

preprocess_minimum_passing_read_length: 51

Require this fraction of each nucleotide per sequence to eliminate low complexity reads.

preprocess_minimum_base_frequency: 0.05

Contamination reference sequences in the form of nucleotide FASTA files can be provided and filtered from the reads using the following parameters.

If ‘rRNA’ is defined, it will be added back to metagenomes but not to metatranscriptomes. Additional references can be added arbitrarily, such as::

contaminant_references:
    rRNA: /database_dir/silva_rfam_all_rRNAs.fa
    phiX: /database_dir/phiX174_virus.fa

Don’t look for indels longer than this:

contaminant_max_indel: 20

Fraction of max alignment score required to keep a site:

contaminant_min_ratio: 0.65

mapping kmer length; range 8-15; longer is faster but uses more memory; shorter is more sensitive:

contaminant_kmer_length: 12

Minimum number of seed hits required for candidate sites:

contaminant_minimum_hits: 1

Set behavior on ambiguously-mapped reads (with multiple top-scoring mapping locations):

contaminant_ambiguous: best

For host decontamination we suggest the following genomes, where contaminants and low complexity regions were masked.