Using de Bruijn Graphs for Short-read Assembly
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Using de Bruijn Graphs for Short-read Assembly

About the Book

About the Author

Homolog_us
Homolog_us

Homolog.us blog provides cutting-edge information on bioinformatics, transcriptomics and computational biology. Our tutorials provide simple introduction for biologists on algorithms related to assembly and analysis of next-generation sequencing. Here we are publishing the tutorials in book format for easy access.

Table of Contents

  • 1. Introduction
    • Assembly procedure requires an understanding of sequencing technologies, algorithms and statistics
      • Sequencing technologies
      • Algorithms
      • Statistics
    • Why should biologists learn about assembly algorithms?
      • Software tools change, but the algorithms remain more stable
      • Knowledge of algorithms helps in experimental design
      • Understanding algorithms helps in purchasing adequate computing hardware
      • Improving assembly quality and interpreting results better
      • RNA-seq in non-model organisms
      • Using NGS technologies to solve novel problems
    • Description of this book
      • Chapters
      • Algebraic notations are avoided as much as possible
      • Living electronic book
      • Core concepts are reinforced through repetition
    • Regarding the exercises
      • Code - Pandora’s Toolbox for Bioinformatics
      • Data - E. coli and Electrophorus
    • Other online resources for learning
      • Introductory resources
      • Intermediate Resources
      • Advanced resources
    • Acknowledgements for this book
    • Further readings
  • 2. The Genome Assembly Problem
    • Shotgun sequencing and assembly of genomes
    • Is the problem solvable?
      • Lander-Waterman statistics for read length and coverage
      • Log(N) estimate for random sequences
      • Ukkonen’s condition
      • Optimum read length and coverage in presence of repeats
      • Reconstruction with paired end read
      • Summary
    • Greedy, overlap-layout-consensus and de Bruijn graph-based algorithms
      • Definitions
      • Greedy algorithm
      • Overlap-layout-consensus algorithm
      • de Bruijn graph-based algorithm
    • A short historical overview of using de Bruijn graphs in genome assembly
    • Advantages and disadvantages of using de Bruijn graphs for assembly
    • Further readings
  • 3. De Bruijn Graph of the Genome and a Simple Assembler
    • De Bruijn Graph of a known genome
      • De Bruijn graph of a small sequence
      • Double-stranded nature of the genome
      • De Bruijn graph in repetitive regions
    • Properties of de Bruijn Graph
      • The graph structure is unique for a given set of kmers
      • Irreversible
      • Impact of changing kmer size
      • Generalized definition of de Bruijn graph
    • Viewing de Bruijn graphs
      • Graphview
      • Online method - Alex Hadik
      • Ray cloud browser
      • FASTG Viewer BANDAGE
    • De Bruijn Graph of the E. coli genome
    • Genome assembly from de Bruijn graphs in the absense of noise
      • Relationship between de Bruijn graph of short reads and the underlying genome
      • Loss of read coherence during de Bruijn graph construction
      • Varying k-mer sizes to compensate for the loss of read coherence
      • Memory requirement and k-mer distribution of perfect library
    • Summary
    • Further readings
  • 4. Experimental Considerations
    • Evolution of sequencing technologies
      • Sanger sequencing
      • 454 pyrosequencing
      • Illumina dye sequencing
      • ABI SOLiD sequencing
      • Ion semiconductor sequencing
      • PacBio single molecule real time sequencing
    • Sources of artifacts
      • Random errors - substitutions and insertion-deletions
      • Homopolymer error
      • Quality drop near 3’ ends of reads
      • Coverage bias in AT-rich or GC-rich regions
      • Distance between the read pairs is variable in mate pairs
      • Mate pair inversion
      • Duplication due to PCR amplification
      • Inherent noise of PacBio reads
      • Diploid genome
    • Paired-end and mate-pair reads
      • Paired end
      • Mate pairs
      • Insert size
    • Data formats
      • FASTA and FASTQ
      • BAM and SAM Alignments
      • CIGAR
    • Detailed description of ABI SOLiD color space data
      • How to convert sequences to color space?
      • How do we compute reverse complement in color space?
      • Simple sequences
      • SNP
      • Advantage of color space
      • Disadvantage
      • Is conversion to nucleotide space prudent?
      • Pseudo-basespace
      • Error correction
      • de Bruijn Graph of SOLiD Reads
    • Summary of what we learned so far
    • Further readings
  • 5. Genome Assembly from Noisy Reads with Uneven Coverage
    • Errors lead to high RAM usage for de Bruijn graph-based assemblers
    • Impact of sequencing errors on the structure of de Bruijn graphs
      • Tips
      • Bubbles
      • Crosslinks
      • Tips and bubbles can appear due to read errors or polymorphism
    • Impact of uneven coverage
    • Full conceptual picture of short read assembly using de Bruijn graph
      • Effect of changing the coverage cutoff parameter
      • Effect of changing the k-mer size
    • Further readings
  • 6. Assembling Transcriptomes, Metagenomes and Heterozygous Genomes
    • General approach for solving complex assembly problems related to short reads
    • Using de Bruijn graphs to assembles heterozygous genomes
      • Impact of haplotype differences on de Bruijn Graph structure
      • Coverage
      • Assembly method
      • Separating phases after assembly
    • Using de Bruijn graphs for transcriptome (RNA-seq) assembly
      • De Bruijn graph structure of transcriptomic libraries
      • K-mer coverage
      • Assembly method
    • Using de Bruijn graphs for metagenome assembly
      • De Bruijn graph structure for metagenomic libraries
      • Coverage
      • Assembly method
    • Further readings
  • 7. Faster, Better and Cheaper
    • Computer science concepts for advanced work
      • Architecture of modern computer
      • Conference analogy
      • Processing Elements - CPU, GPU and FPGA
      • Algorithm and data structure
      • Disk-based algorithms and Hadoop
      • Parallel code, compare-and-swap, shared memory and queues
      • Hashing-related concepts
      • Alignment-related concepts
      • Other algorithmic concepts related to bioinformatics
    • Data structures for efficient storage and processing of de-Bruijn graphs
      • An elaborate data structure for de Bruijn graph
      • Using simplified k-mers - ABySS
      • Using edge-based data structure - Conway and Bromage
      • Using Sparse de Bruijn Graph - Sparse-assembler
      • Using Bloom filter - Minia
      • Using Perfect hash - Meraculous
      • Using Minimizer - BCALM
      • Using Succint Data Structure
    • Efficient algorithms for counting k-mers
      • Meryl
      • Suffix array - Tallymer
      • Bloom filter - BFcounter and scTurtle
      • Lock-free hash table - Jellyfish
      • Disk-based - DSK, KMC and KAnalyze
      • Minimizer-based - MSPKmerCounter, KMC2 and DSK2
      • Approximate counting - khmer
    • Efficient algorithms for read error correction
    • Improving assembly by changing k-mer size for de Bruin graph
      • Optimal k-mer-size for dBG construction
      • Merging read pairs to increase effective read length
      • Combining assemblies from multiple k-mers - IDBA and SPAdes
      • Variable order k-mer
      • String Graph Assembler for Short Reads
    • Scaffolding
      • Hierarchical scaffolding - SOAPdenovo
      • Rectangular Graph - SPAdes
    • Repeat resolution
      • SOAPdenovo
    • SPAdes
      • Hyperconnected k-mers
    • Further readings
  • 8. In Depth Discussion of Three de Bruijn Graph-based Assemblers
    • Where to get them
    • Genome assembler - SOAPdenovo
      • History
      • How to run
      • Features
      • Details of algorithm
      • Details of code
      • SOAPdenovo-trans Transcriptome Assembler
    • Genome Assembler - SPAdes
      • History
      • How to run
      • Features of SPAdes
      • Details of algorithm
      • Details of code
    • Transcriptome assembler - Trinity
      • History
      • How to run
      • Details of algorithm
      • Details of code
    • Further readings
  • 9. References
    • 1. Pre-NGS genome assemblers
      • Base-calling and error detection
      • Assemblers
    • 2. NGS genome assemblers
      • non de Bruijn, k-mer based
      • de Bruijn graph-based assemblers
      • Applications
      • Comparison
    • 3. Exomes, transcriptomes, metagenomes and highly polymorphic genomes
      • Transcriptome assemblers
      • Metagenomes
      • Polymorphic genomes
      • Targeted assembly
    • 4. Faster, better, cheaper
      • k-mer counting
      • Storage
      • Error correction
      • Hadoop
      • Hardware accelerators
      • String graph assembler
      • Scaffolding
      • Repeats
    • 6. Reviews and forecasts

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