From 334625db8baab6c26d45a34beb24770d8a13755d Mon Sep 17 00:00:00 2001 From: Tom Harrop Date: Mon, 4 Nov 2024 11:57:35 +1100 Subject: [PATCH 1/3] mvp for trimming --- tools/dorado/dorado_trimming.xml | 57 ++++++++++++++++++++++++++++++++ 1 file changed, 57 insertions(+) create mode 100644 tools/dorado/dorado_trimming.xml diff --git a/tools/dorado/dorado_trimming.xml b/tools/dorado/dorado_trimming.xml new file mode 100644 index 00000000..0b70aba6 --- /dev/null +++ b/tools/dorado/dorado_trimming.xml @@ -0,0 +1,57 @@ + + for removing adapter and/or primer sequences from the beginning and end of Nanopore DNA reads + + macros.xml + + + + trimmed.bam + +&& + +dorado summary +trimmed.bam +> summary.tsv + + + ]]> + + + + + + + + + + + + + + + + + + + + + + + + + + + From 0fcfbf2accb6dbb7318d7ce35a921c2875beb044 Mon Sep 17 00:00:00 2001 From: Tom Harrop Date: Mon, 4 Nov 2024 13:54:28 +1100 Subject: [PATCH 2/3] more tests --- tools/dorado/dorado_trimming.xml | 16 ++++++++++++++++ 1 file changed, 16 insertions(+) diff --git a/tools/dorado/dorado_trimming.xml b/tools/dorado/dorado_trimming.xml index 0b70aba6..bf01a1c0 100644 --- a/tools/dorado/dorado_trimming.xml +++ b/tools/dorado/dorado_trimming.xml @@ -12,9 +12,13 @@ ln -s '$reads' ./reads && dorado trim +--threads "\${GALAXY_SLOTS}" #if $no_trim_primers --no-trim-primers #end if +#if $primer_sequences + --primer-sequences '$primer_sequences' +#end if reads > trimmed.bam @@ -49,6 +53,18 @@ trimmed.bam + + + + + + + + + + + + Date: Mon, 4 Nov 2024 14:14:33 +1100 Subject: [PATCH 3/3] tests --- tools/dorado/dorado_trimming.xml | 61 +++++++++++++----- .../FAL00375_473bf0ed_0.ten_reads.bam | Bin 0 -> 60959 bytes .../dorado/test-data/custom_primers.fasta.gz | Bin 0 -> 51 bytes .../test-data/dorado_trimming_test1.bam | Bin 0 -> 60520 bytes .../test-data/dorado_trimming_test1.tsv | 11 ++++ .../test-data/dorado_trimming_test2.bam | Bin 0 -> 60621 bytes .../test-data/dorado_trimming_test2.tsv | 11 ++++ .../test-data/dorado_trimming_test3.bam | Bin 0 -> 787 bytes .../test-data/dorado_trimming_test3.tsv | 2 + .../lsk109_single_read.fastqsanger.gz | Bin 0 -> 589 bytes 10 files changed, 69 insertions(+), 16 deletions(-) create mode 100644 tools/dorado/test-data/FAL00375_473bf0ed_0.ten_reads.bam create mode 100644 tools/dorado/test-data/custom_primers.fasta.gz create mode 100644 tools/dorado/test-data/dorado_trimming_test1.bam create mode 100644 tools/dorado/test-data/dorado_trimming_test1.tsv create mode 100644 tools/dorado/test-data/dorado_trimming_test2.bam create mode 100644 tools/dorado/test-data/dorado_trimming_test2.tsv create mode 100644 tools/dorado/test-data/dorado_trimming_test3.bam create mode 100644 tools/dorado/test-data/dorado_trimming_test3.tsv create mode 100644 tools/dorado/test-data/lsk109_single_read.fastqsanger.gz diff --git a/tools/dorado/dorado_trimming.xml b/tools/dorado/dorado_trimming.xml index bf01a1c0..d4c1c778 100644 --- a/tools/dorado/dorado_trimming.xml +++ b/tools/dorado/dorado_trimming.xml @@ -1,5 +1,5 @@ - for removing adapter and/or primer sequences from the beginning and end of Nanopore DNA reads + for Oxford Nanopore (ONT) DNA reads macros.xml @@ -12,6 +12,7 @@ ln -s '$reads' ./reads && dorado trim +--verbose --threads "\${GALAXY_SLOTS}" #if $no_trim_primers --no-trim-primers @@ -42,32 +43,60 @@ trimmed.bam - - - - - - - - - - + + - - + + - - + + `__ basecaller. + +This tool scans existing, basecalled datasets for adapter and/or primer +sequences at either end, and trims any such found sequences. + +**If you have raw (un-basecalled) data, you can trim them during +basecalling with the Dorado tool on Galaxy**. + +Note that if you intend to demultiplex the reads later, trimming +adapters and primers may result in some portions of the flanking regions +of the barcodes being removed, which could interfere with correct +demultiplexing. + +The **Don't trim primers** option can be used to prevent the trimming of +primer sequences. In this case only adapter sequences will be trimmed. + +The output of will always be unaligned records, regardless of whether +the input is aligned/sorted or not. + +Custom primer trimming +---------------------- + +The software automatically searches for primer sequences used in Oxford +Nanopore kits. However, you can specify an alternative set of primer +sequences to search by adding a FASTA file of primer sequences in the +**Custom primer sequences** option. The record names of the sequences do +not matter. Note that if you use this option the normal primer sequences +built-in to the dorado software will not be searched for. + +RNA adapter trimming +-------------------- + +Adapters for RNA002 and RNA004 kits are automatically trimmed during +basecalling. 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