Address of the bookmark: http://bioinformatics.oxfordjournals.org/content/early/2014/02/12/bioinformatics.btu078.full.pdf

With Single Molecule, Real-Time (SMRT) Sequencing, you can access structural variations having a broad range of sizes, types, and GC content with the ability to:

• Uncover missing heritability linked to structural variation
• Unambiguously identify genomic context and variant breakpoints at the sequence level to unravel the genetic etiology of disease
• Resolve structural variation across the complete size spectrum with basepair resolution

Following are the SV tools, which can assist you to achieve your goal.

Sniffles: Structural variation caller using third generation sequencing

Sniffles is a structural variation caller using third generation sequencing (PacBio or Oxford Nanopore). It detects all types of SVs using evidence from split-read alignments, high-mismatch regions, and coverage analysis. Please note the current version of Sniffles requires sorted output from BWA-MEM (use -M and -x parameter) or NGM-LR with the optional SAM attributes enabled! 

More at https://github.com/fritzsedlazeck/Sniffles

MultiBreak-SV: It identifies structural variants from next-generation paired end data, third-generation long read data, or data from a combination of sequencing platforms.

There are two pieces of software in this release: (1) a pre-processor that takes machineformat (.m5) BLASR files, and (2) MultiBreak-SV. For installation and usage instructions, see doc/MultiBreakSV-Manual.txt.

More at https://github.com/raphael-group/multibreak-sv

Parliament: A Structural Variation Tool. Why ask a single sv-detection approach to find every variant when you can have a parliament of tools deciding?

Publication about the algorithm and “…the first long-read characterization of structural variation in a diploid human personal genome…” (HS1011) - “Assessing structural variation in a personal genome—towards a human reference diploid genome”

More at https://sourceforge.net/projects/parliamentsv/

https://www.dnanexus.com/papers/Parliament_Info_Sheet.pdf

PBHoney: the structural variation discovery tool 

PBHoney is an implementation of two variant-identification approaches designed to exploit the high mappability of long reads (i.e., greater than 10,000 bp). PBHoney considers both intra-read discordance and soft-clipped tails of long reads to identify structural variants.

Read The Paper http://www.biomedcentral.com/1471-2105/15/180/abstract

More at https://sourceforge.net/projects/pb-jelly/

SMRT-SV: Structural variant and indel caller for PacBio reads

Structural variant (SV) and indel caller for PacBio reads based on methods from Chaisson et al. 2014.

SMRT-SV provides an official software package for tools described in Chaisson et al. 2014 and adds several key features including the following.

• Unified variant calling user interface with built-in cluster compute support
• Small indel calling (2-49 bp)
• Improved inversion calling (screenInversions)
• Quality metric for SV calls based on number of local assemblies supporting each call
• Higher sensitivity for SV calls using tiled local assemblies across the entire genome instead of "signature" regions
• Genotyping of SVs with Illumina paired-end reads from WGS samples

More at https://github.com/EichlerLab/pacbio_variant_caller

Metagenomic simulators are a way of dealing with this issue; they produce artificial sequencing datasets in which both the composition of the microorganisms and the expected results are known. Researchers can then test a pipeline on such a controlled dataset and see precisely how well it functions. However, as metagenomic studies have become larger, more diverse, and more complex, simply generating artificial reads is no longer sufficient. The simulators now have to replicate community variation, sequencing errors, different sequencing platforms, massive data volumes, and even laboratory-based biases.

MIDASim (2024) was designed with a particular emphasis on microbial community dynamics. Unlike previous tools which regarded each simulated sample as individual, MIDASim is able to efficiently generate realistic multi-sample microbiome datasets, covering the cases where changes over time are being tracked. This feature is particularly useful when investigating how microbial communities vary between people, different experimental setups, or at various times. By overcoming the computational limitations of earlier simulation methods, MIDASim has made it easier to carry out large and complex microbiome simulation studies.

A major challenge is scale. Since modern metagenomic projects can involve millions or even billions of sequencing reads, generating such large synthetic datasets can become a significant computational problem. In order to tackle this issue, Izzy (2025) was developed. As a high-throughput metagenomic read simulator, Izzy is designed with speed in mind while still reproducing the typical patterns of sequencing errors. The people who developed it stated that it can be as much as 60 times faster than earlier tools, which makes large-scale simulations far more practical. Now, researchers can begin to ask not how long a simulation will take but rather what size dataset they want to test.

At the same time, the sequencing technology became more diverse. Researchers were not any longer restricted to using Illumina short reads since Oxford Nanopore and PacBio long-read sequencing also became important in the field of metagenomic research. In order to address these new requirements, MeSS (Metagenomic Sequence Simulator, 2025) was developed. The programme is based on a Snakemake framework and is therefore compatible with the Illumina, Oxford Nanopore and PacBio platforms. MeSS has also been designed to operate efficiently and to use less memory than earlier tools; it provides ready-made templates for the microbiomes of different sites in the human body, thus giving researchers a simple means of beginning to generate realistic simulated communities.

The simulation problem became even more specialized. Suppose that sequencing is not based on uniform sampling of DNA? In the case of targeted sequencing and hybridization-capture experiments, particular sequences are deliberately enriched, and the probability of capturing a target depends on both the probe and the target sequence. A standard model based on uniform sampling might fail to take into account this important feature of real experiments. That is why RAmpSim has been designed specifically to handle simulations for capture-based and targeted sequencing. Instead of depending solely on the assumption of uniform sampling, it includes a thermodynamic nearest-neighbor energy model to represent probe–target interactions and sequence-dependent enrichment. As a result, it is especially suitable for applications such as hybridization capture in metagenomics, where the efficiency of recovering a sequence can depend strongly on how it relates to the capture probes. RAmpSim thus reflects a wider trend towards simulations that aim not only to reproduce sequencing output but also key aspects of the experimental process itself.

MIDASim, Izzy, MeSS and RAmpSim all demonstrate how fast metagenomic simulation is evolving. MIDASim is able to deal with realistic variation within a large number of and changing microbial communities. Izzy overcomes the problem of generating very large datasets. MeSS provides support for a number of sequencing technologies in an efficient and repeatable manner. RAmpSim increases the experimental realism for capture-based sequencing. Although each of these tools addresses a separate issue, they all contribute to advancing the field.

Modern metagenomic simulation therefore aims not only at producing artificial FASTQ files. Researchers nowadays desire simulated datasets that reflect community changes, incorporate the characteristics of the sequencing platform, include real error patterns, take into account computational scale, and account for experimental biases. As metagenomic workflows become more advanced, the simulators have to keep up with this development. The tools are now going beyond that of simple data generators; they produce controlled versions of complex metagenomic experiments and thus provide researchers with something that ordinary sequencing data seldom does: a dataset in which the answer is known before any analysis takes place.

• Assembly of nanopore reads using Canu.
• Polish canu contigs using racon (optional).
• Map a paired-end Illumina dataset onto the contigs obtained in the previous steps using BWA mem.
• Perform correction of contigs using pilon and the Illumina dataset.

Address of the bookmark: https://github.com/nanoporetech/ont-assembly-polish

• dnaPipeTE is developped by Clément Goubert, Laurent Modolo and the TREEP team of the LBBE: http://lbbe.univ-lyon1.fr/-Equipe-Elements-transposables-.html?lang=en

• You can find the original publication in GBE here: https://academic.oup.com/gbe/article/7/4/1192/533768

output examples of quantification and TE landscape (relative age) produced by dnaPipeTE

Address of the bookmark: https://github.com/clemgoub/dnaPipeTE

Ra is in development since 2014 in the form of several separate components that used to be run individually.
This project aims to ease the usage of Ra by integrating it into a complete de novo assembly tool.

Unlike other state-of-the-art assemblers, Ra does not have an error correction step. Instead, it relies on detecting overlaps using a very sensitive and specific overlapper ("graphmap -w owler", https://github.com/isovic/graphmap) and constructing and reducing an overlap graph (Ra layout, https://github.com/mariokostelac/ra).

Address of the bookmark: https://github.com/mariokostelac/ra-integrate/

Using AlignGraph

AlignGraph --read1 reads_1.fa --read2 reads_2.fa --contig contigs.fa --genome genome.fa --distanceLow distanceLow --distanceHigh distancehigh --extendedContig extendedContigs.fa --remainingContig remainingContigs.fa [--kMer k --insertVariation insertVariation --coverage coverage --part p --fastMap --ratioCheck --iterativeMap --misassemblyRemoval --resume]

Address of the bookmark: https://github.com/baoe/AlignGraph

This script has been modified by others to convert InDels as well, e.g. this by David Eccles

./vcf2fq.pl -f <input.fasta> <all-site.vcf> > <output.fastq>

https://github.com/gringer/bioinfscripts/blob/master/vcf2fq.pl

https://github.com/lh3/samtools/blob/master/bcftools/vcfutils.pl