{ "info": { "author": "Jose Fernandez Navarro", "author_email": "jose.fernandez.navarro@scilifelab.se", "bugtrack_url": null, "classifiers": [ "Development Status :: 5 - Production/Stable", "Environment :: Console", "Intended Audience :: Science/Research", "License :: OSI Approved :: MIT License", "Operating System :: MacOS", "Operating System :: Unix", "Programming Language :: Python :: 3.6", "Programming Language :: Python :: 3.7", "Topic :: Scientific/Engineering :: Bio-Informatics", "Topic :: Software Development" ], "description": "The ST Pipeline contains the tools and scripts needed to process \nand analyze the raw files generated with the Spatial Transcriptomics \nmethod in FASTQ format to generated datasets for down-stream analysis. \nThe ST pipeline can also be used to process single cell data as \nlong as a file with barcodes identifying each cell is provided.\nThe ST Pipeline can also process RNA-Seq datasets generated with \nor without UMIs. \n\nThe ST Pipeline has been optimized for speed, robustness and it is very \neasy to use with many parameters to adjust all the settings.\n\nThe following files/parameters are required:\n\n- FASTQ files (Read 1 containing the spatial information and the UMI \n and read 2 containing the genomic sequence) \n- A genome index generated with STAR \n- An annotation file in GTF or GFF format (optional)\n- The file containing the barcodes and array coordinates \n (look at the folder \"ids\" and chose the correct one). \n Basically this file contains 3 columns (BARCODE, X and Y), \n so if you provide this file with barcodes identinfying cells (for example), \n the ST pipeline can be used for single cell data.\n This file is optional too. \n- A name for the dataset\n\nThe ST pipeline has multiple parameters mostly related to trimming, \nmapping and annotation but generally the default values are good enough. \nYou can see a full description of the parameters \ntyping \"st_pipeline_run.py --help\" after you have installed the ST pipeline.\n\nThe input FASTQ files can be given in gzip/bzip format as well. \n\nBasically what the ST pipeline does is:\n\n- Quality trimming (read 1 and read 2):\n\t- Remove low quality bases\n\t- Sanity check (reads same length, reads order, etc..)\n\t- Check quality UMI (if provided)\n\t- Remove artifacts (PolyT, PolyA, PolyG, PolyN and PolyC) of user defined length\n\t- Check for AT and GC content\n\t- Discard reads with a minimum number of bases of that failed any of the checks above\n- Contamimant filter e.x. rRNA genome (Optional)\n- Mapping with STAR (only read 2)\n- Demultiplexing with [Taggd](https://github.com/SpatialTranscriptomicsResearch/taggd) (only read 1)\n- Keep reads (read 2) that contain a valid barcode and are correctly mapped\n- Annotate the reads with htseq-count (optional)\n- Group annotated reads by barcode(spot position) and gene to get a read count\n- In the grouping/counting only unique molecules (UMIs) are kept. \n\nYou can see a graphical more 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