Supported data types
Sequencing data
Spl-IsoQuant support all kinds of long RNA data:
- PacBio CCS
- ONT dRNA / ONT cDNA
- Assembled / corrected transcript sequences
Reads must be provided in FASTQ/FASTA format (can be gzipped) or unmapped BAM format.
If you have already aligned your reads to the reference genome, simply provide sorted and indexed BAM files.
Spl-IsoQuant supports the following protocols:
- 10x 3' v3 single-cell;
- 10x 3' Visium spatial data;
- 10x Visium HD;
- Curio Biosciences spatial data;
- Stereo-seq spatial data;
- Any other single-cell or spatial protocol with barcode and UMI sequences (see more about custom molecule description).
Supported reference data
Reference genome should be provided in multi-FASTA format (can be gzipped). The reference genome is mandatory even when BAM files are provided.
Reference gene annotation is also mandatory for single-cell / spatial analysis
It should be provided in GFF/GTF format (can be gzipped).
It will be converted to gffutils database. Information on converted databases will be stored in your ~/.config/Spl-IsoQuant/db_config.json to increase speed of future runs. You can also provide gffutils database manually.
Make sure that chromosome/scaffold names are identical in FASTA file and gene annotation.
Note that gffutils databases may not work correctly on NFS shares. It is possible to set a designated folder for
the database with --genedb_output (different from the output directory).
Pre-constructed aligner index can also be provided to reduce mapping time.