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Fix multiplexed isoseq example (#60)
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Co-authored-by: Jessica Mattick <[email protected]>
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jmattick and Jessica Mattick authored Jun 27, 2024
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### Single sample

This is a toy dataset consisting of ~80k segmented reads (S-reads) from a Kinnex full-length RNA library. The original HiFi reads have already been segmented using Skera, so we can directly go into primer removal using lima and such.

# Download toy S-read dataset
$ wget https://downloads.pacbcloud.com/public/dataset/IsoSeq_sandbox/human_80k_Sreads.segmented.bam

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flnc.report.csv

$ isoseq cluster2 flnc.bam transcripts.bam


$ ls transcripts*
transcripts.bam
transcripts.bam.pbi
transcripts.cluster_report.csv

### Multiplexed samples

This is a 12-plex regular Iso-Seq (non-Kinex) run on Sequel II system consisting of ~3 million HiFi reads.
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$ wget https://downloads.pacbcloud.com/public/dataset/Kinnex-full-length-RNA/REF-primers/IsoSeq_v2_primers_12.fasta

# Demux and primer removal
$ lima --isoseq --peek-guess m64307e_230628_025302.hifi_reads.bam IsoSeq_v2_primers_12.fasta output.bam
$ lima --overwrite-biosample-names --isoseq --peek-guess m64307e_230628_025302.hifi_reads.bam IsoSeq_v2_primers_12.fasta output.bam

# Combine inputs
$ ls output.IsoSeqX*bam > all.fofn
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