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Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.

Appendix A

Tables of Computational Tools

TABLE A-1 State-of-the-Art Nanopore Basecalling Tools with Principle of Analysis, Input, and Output Along with Resources

ToolPrinciple of AnalysisTool InputTool OutputResource
AlbacoreRNNFAST5FASTA/FASTQhttps://community.Nanoporetech.com
GuppyRNNFAST5FASTA/FASTQhttps://community.Nanoporetech.com
MinKNOW
MetrichorRNNFAST5FASTA/FASTQhttps://metrichor.com
NanocallHMMFAST5FASTAhttps://github.com/mateidavid/nanocall
DeepNanoRNNFAST5FASTAhttps://github.com/jeammimi/deepnano
NanonetRNNFAST5FASTAhttps://github.com/ProgramFiles/nanonet
basecRAWllerhttps://www.osti.gov/biblio/1572483
ChironCNNs, RNNs, and CTC decoderFAST5FASTA/FASTQhttps://github.com/haotianteng/chiron
ScrappieRNNFAST5FASTA/SAMhttps://github.com/Nanoporetech/scrappie
BonitoQuartzNet13(multiple TCSConv-BNReLU)FAST5FASTQ, SAM, BAM, CRAMhttps://github.com/Nanoporetech/bonito
Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.
ToolPrinciple of AnalysisTool InputTool OutputResource
URNanoRefined U-net model for signal segmentation and basecallingFAST5FASTA/FASTQhttps://github.com/yaozhong/URnano
CausalcallTCNFAST5FASTA/FASTQhttps://github.com/scutbioinformatic/causalcall
Heron and OspreyCNN with dynamic poolingFAST5FASTAhttps://github.com/fmfi-compbio/heron
https://github.com/fmfi-compbio/osprey
DeepNano-coralEnhanced version of Bonito-CNN that has factorization of a full convolution into smaller operationsFAST5FASTAhttps://github.com/fmfi-compbio/coral-training
DeepNano-blitzBi-directional RNNFAST5FASTAhttps://github.com/fmfi-compbio/deepnano-blitz
BRAWLBinarization of Chiron basecaller (CNN+RNN+CTC)FAST5FASTA/FASTQhttps://github.com/haotianteng/Chiron
SACallConvolution layers, transformer self-attention layers and a CTC decoderFAST5FASTA/FASTQhttps://github.com/huangnengCSU/SACall-basecaller
CATCallerConvolution-augmented transformer architectureFAST5FASTA/FASTQhttps://github.com/biomed-AI/CATCaller
HalcyonCNN module and RNN-based encoder and decoderFAST5FASTAhttps://github.com/relastle/halcyon
ToolPrinciple of AnalysisTool InputTool OutputResource
RavventEncoder-decoder architecture with attention mechanism and LSTMs as RNNsFAST5FASTAhttps://github.com/adamnapieralski/ravventbasecaller
DoradoBased on libtorch, the C++ API for PyTorchPOD5HTS formathttps://github.com/nanoporetech/dorado

NOTE: CNN = convolutional neural network; CTC = connectionist temporal classification decoder; TCN = temporal convolutional network; LSTM = long short-term memory network; API = application programming interface. Other abbreviations are defined in the Front Matter.

Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.

TABLE A-2 Various Quality Control Tools Used with Nanopore Sequencing Data

QC ToolPrinciple of AnalysisInputOutputResource
PoretoolsOpen-source software and is written in PythonFAST5 filesTables & Graphs (PNG & PDF)https://github.com/arq5x/poretools
poReCross-platform tool scripted in R languageFAST5 filesTables & Graphshttps://github.com/mw55309/poRe_docs
NanoOKAlignment-based QC tool, written in Java with supporting R scriptsFAST5 filesPDF reporthttps://github.com/TGAC/NanoOK
NanoPackLong-read QC tool written in Python languageFASTQ, sorted BAM, & sequencing summary filesHTML, PDF, PNG & JPGhttps://github.com/wdecoster/nanopack
ToulligQCQC tool for guppy basecalled files, written in Python 3Sequencing summary filesHTMLhttps://github.com/GenomicParisCentre/toulligQC
MinIONQCNon-interactive tool written in RSequencing summary filePNG & YAML formathttps://github.com/roblanf/minion_qc
PycoQCInteractive QC tool written in Python languageSequencing summary fileHTMLhttps://github.com/a-slide/pycoQC
NanoRA cross-platform R packageFAST5 file or sequencing summary fileTSV file or HTML filehttps://github.com/davidebolo1993/NanoR
PyPoreOpen-source interactive software written in Python 2.7FAST5 filesHTMLhttps://github.com/rsemeraro/PyPore
LongQCAutomated QC tool written in Python 3FASTA, FASTQ, or a PacBio BAM fileJSON and HTMLhttps://github.com/yfukasawa/LongQC
BoardIONWeb application based on RFAST5 file or sequencing summary fileHTMLhttps://github.com/institut-degenomique/BoardION
IONiseRR packageFAST5R plotshttps://www.bioconductor.org/packages/release/bioc/html/IONiseR.html
Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.

TABLE A-3 ONT Assemblers for Long-Reads

Name of the toolPrinciple of analysisInputOutputResource
MaSuRCADe Bruijn graph and overlap-layout-consensusFASTA/FASTQFASTAhttps://github.com/alekseyzimin/masurca
CanuDetection of overlaps in high-noise sequences using MinHash Alignment ProcessFASTA/FASTQ, uncompressed or compressed in gzip, bzip2, or xzFASTAhttps://github.com/marbl/canu
UnicyclerMiniasm+Racon pipeline through a short-read-first hybrid assemblyFASTQFASTAhttps://github.com/rrwick/Unicycler
HINGEVariation of the greedy algorithm, called hinging without a preassembly or read correction stepFASTAFASTAhttps://github.com/HingeAssembler/HINGE
FlyeRepeat graphs from the analysis of random paths in an unknown repeat graph, called disjointigsFASTQ or FASTA format, uncompressed or compressed with gzipFASTAhttps://github.com/fenderglass/Flye
ShastaRun-length representation of the read sequenceFASTA/FASTQFASTA/GFAhttps://github.com/chanzuckerberg/shasta
Wtdbg2FBG to long noisy reads assemblyFASTA/FASTQFASTAhttps://github.com/ruanjue/wtdbg2

NOTE: FBG = fuzzy De Bruijn graph.

Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.

TABLE A-4 Aligners for RNA-Seq Data

ToolPrinciple of analysisInputOutputSource
HISATMapping next-generation sequencing reads (both DNA and RNA) to a population of human genomes as well as to a single reference genomeFASTA/FASTQ/SEQSAMhttps://github.com/DaehwanKimLab/hisat2
Minimap2Overlapping read-to-read sequences with k-mersFASTA/FASTQSAM/PAFhttps://github.com/lh3/minimap2
DARTSeparation of sequence into segments to replace the seed extension stepFASTA/FASTQSAM/BAMhttps://github.com/hsinnan75/Dart
Magic-BLASTSelection of the best scoring alignment from sequences found in the NCBIFASTASAM/BAMhttps://github.com/ncbi/magicblast/tree/master/magicblast-tools
GraphMap2Subdivision of k-mer similarityFASTA/FASTQSAMhttps://github.com/lbcb-sci/graphmap2
deSALT`De Bruijn graph-based spliced aligner for long-readsFASTA/FASTQSAMhttps://github.com/ydLiuHIT/deSALT
uLTRAAlignment of reads to a genome through guided annotation of exonsFASTA/FASTQSAMhttps://github.com/ksahlin/ultra
mapAlignMap long-reads against a reference genome or a prebuilt index fileFASTA/FASTQSAMhttps://github.com/yw575/mapAlign

NOTE: NCBI = National Center for Biotechnology Information

Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.

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Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.
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Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.
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Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.
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Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.
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Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.
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Suggested Citation: "Appendix A: Tables of Computational Tools." National Academies of Sciences, Engineering, and Medicine. 2024. Charting a Future for Sequencing RNA and Its Modifications: A New Era for Biology and Medicine. Washington, DC: The National Academies Press. doi: 10.17226/27165.
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Next Chapter: Appendix B: Public Meeting Agendas
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