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Assembly ā€“ before and after
Lex Nederbragt
lex.nederbragt@ibv.uio.no
@lexnederbragt
A warning
The list is by no means complete
Nor do we have experience with all the programs mentioned
Sample DNA Reads
Genome
assembly
Sequencing AssemblyDNA isolation
QC QCQC
Reads
Genome
assembly
Assembly
QC
Fastqc
Prinseq
Many othersā€¦
www.nipgr.res.in/ngsqctoolkit.html
preqc (sga)
http://arxiv.org/abs/1307.8026
Reads
Genome
assembly
Assembly
Grooming
Format conversion
http://en.wikipedia.org/wiki/FASTQ_format
Fastq format hell
Adapter/quality trimming
http://www.biostars.org/p/53528/
Celera assembler
Overlap based trimming
Fastx Toolkit
Seqtk
PrinSeq
NGS QC Toolkit
Trimmomatic
BioPieces
Cutadapt
ā€¦
ā€¦
Mate pair splitting and orientation
150 ā€“ 600 bases
Illumina paired end reads
2 ā€“ 40 kilobases
Illumina mate pair reads
2 ā€“ 40 kilobases
454 mate pair reads
linker
Mate pair splitting and orientation
Illumina paired end reads
Illumina mate pair reads
454 mate pair reads
linker
junctionjunction
+ +
paired end reads
ā€˜contaminationā€™
Mate pair splitting and orientation
Illumina paired end reads
Illumina mate pair reads
454 mate pair reads
linker
junctionjunction
+ +
paired end reads
ā€˜contaminationā€™
Check what orientation
your assembler expects
for the reads!
Reads
Genome
assembly
Assembly
Preparing
Error-correction
Stand-alone or built into assembler
Merging pairs
List from Torsten Seemanā€™s blog
http://thegenomefactory.blogspot.no/2012/11/tools-to-merge-overlapping-paired-end.html
COPE http://sourceforge.net/projects/coperead/
SeqPrep https://github.com/jstjohn/SeqPrep
FLASH http://www.cbcb.umd.edu/software/flash
fastq-join http://code.google.com/p/ea-utils/wiki/FastqJoin
PANDAseq https://github.com/neufeld/pandaseq
mergePairs.py http://code.google.com/p/standardized-velvet-assembly-report/source/browse/trunk/mergePairs.py
Recent addition
Extend reads
http://140.116.235.124/~tliu/arf-pe/
Digital normalisation
http://arxiv.org/abs/1203.4802
Estimate kmer to use
preqc (SGA)
http://arxiv.org/abs/1307.8026
Reads
Genome
assembly
Assembly
What can the reads tell us about the genome
kmer-based
preqc (SGA)
Kmerspectrumanalyzer
http://arxiv.org/abs/1307.8026
Khmer from Titus
Reads
Genome
assembly
Assembly
This
talk
Reads
Genome
assembly
Assembly
QC
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Assemblathon stats
http://korflab.ucdavis.edu/datasets/Assemblathon/Assemblathon2/Basic_metrics/assembla
thon_stats.pl
OR
https://github.com/lexnederbragt/sequencetools/
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Gap closing
IMAGE2
Correcting bases
Quiver from Pacific Biosciences
Separate scaffolding
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Assembly merging/reconciliation
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Mapped genomic reads
FRCBAM
Mapped transcriptomic reads
Gene finding
Binning
Bacteroides
Proteobacteria
Cyanobacteria
Per-con g read depth
Nederbragt et al, 2010
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Genome browser(s)
IGV
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Comparative measures
Log Average Probability (LAP)
Assembly Likelihood Evaluation (ALE)
See also Howison, Zapata2 and Dunn (2013) Toward a
statistically explicit understanding of de novo sequence
assembly doi: 10.1093/bioinformatics/btt525
Genome
assembly
Comparing to each other
Metrics
Merging
Improvement
Visualization
Validation
Comparing to reference
Reference comparison
Mauve assembly metrics
Review
Too many toolsā€¦
http://seqanswers.com/wiki/Software/list
Too many toolsā€¦
http://wwwdev.ebi.ac.uk/fg/hts_mappers
88 short-read mappers
Embargo!
Benchmarking, anyone?
All-in-one assembly pipeline
doi:10.1186/1471-2105-15-126

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