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CAFU is a Galaxy-based bioinformatics framework for comprehensive assembly and functional annotation of unmapped RNA-seq data from single- and mixed-species samples which integrates plenty of existing NGS analytical tools and our developed programs, and features an easy-to-use interface to manage, manipulate and most importantly, explore large-scale unmapped reads.
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Besides the common process of reads cleansing, reads mapping, unmapped reads generation and novel transcription assembly, CAFU optionally offers the multiple-level evidence analysis of assembled transcripts, the sequence and expression characteristics of assembled transcripts, and the functional exploration of assembled transcripts through gene co-expression analysis and genome-wide association analysis.
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Taking advantages of machine learning (ML) technologies, CAFU also effectively addresses the challenge of classifying species-specific transcripts assembled using unmapped reads from mixed-species samples.
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The CAFU project is hosted on GitHub(https://github.com/cma2015/CAFU) and can be accessed from http://bioinfo.nwafu.edu.cn:4001. The CAFU Docker image is available at https://hub.docker.com/r/malab/cafu.
- Extraction of unmapped reads
- De novo transcript assembly of unmapped reads
- Evidence support of assembled transcripts
- Species assignment of assembled transcripts
- Sequence characterization of assembled transcripts
- Expression profiles of assembled transcripts
- Function annotation of assembled transcripts
- Tutorials for CAFU: https://github.com/cma2015/CAFU/blob/master/Tutorials/User_manual.md
- Test datasets referred in the tutorials for CAFU: https://github.com/cma2015/CAFU/tree/master/Test_data
- In the function Assemble Unmapped Reads, a parameter "Memory" was added for setting the maximum memory to be used by Triniry (1G in default).
- To run the function Species Assignment of Transcripts, users can now use pre-trained or self-trained models. Currently, a pre-trained model was provided by training 20,502 and 137,052 mRNAs annotated in the reference genome of stripe rust pathogen Puccinia striiformis f. sp. tritici (PST-78 v1) and Chinese Spring wheat (IWGSC RefSeq v1.0), respectively.
- The user tutorial was updated to highlight the importance of CPUs, Memory and Swap settings for running CAFU docker.
- A function Remove Contamination was added to remove potential contamination sequences using Deconseq (Schmieder et al., 2011).
- A function Remove Batch Effect was added to remove batch effects using an R package sva (Leek et al., 2012).
- CAFU source codes, web server and Docker image were released for the first time.
- For any bugs/issues, please feel free to leave a message at Github issues. We will try our best to deal with all issues as soon as possible.
- For any suggestions/comments, please send emails to: Siyuan Chen [email protected] or Jingjing Zhai [email protected]
Siyuan Chen, Chengzhi Ren, Jingjing Zhai, Jiantao Yu, Xuyang Zhao, Zelong Li, Ting Zhang, Wenlong Ma, Zhaoxue Han, Chuang Ma, CAFU: A Galaxy framework for exploring unmapped RNA-Seq data. Briefings in Bioinformatics, doi:10.1093/bib/bbz018.