<?xml version='1.0'?><rss version="2.0" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:georss="http://www.georss.org/georss" xmlns:atom="http://www.w3.org/2005/Atom" >
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	<title><![CDATA[BOL: Related items]]></title>
	<link>https://bioinformaticsonline.com/related/34398?offset=0</link>
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	<description><![CDATA[]]></description>
	
	<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/41599/haslr-a-hybrid-assembler-which-uses-both-second-and-third-generation-sequencing-reads</guid>
	<pubDate>Mon, 04 May 2020 02:04:03 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/41599/haslr-a-hybrid-assembler-which-uses-both-second-and-third-generation-sequencing-reads</link>
	<title><![CDATA[HASLR: a hybrid assembler which uses both second and third generation sequencing reads]]></title>
	<description><![CDATA[<p><span>HASLR, a hybrid assembler which uses both second and third generation sequencing reads to efficiently generate accurate genome assemblies. Our experiments show that HASLR is not only the fastest assembler but also the one with the lowest number of misassemblies on all the samples compared to other tested assemblers. Furthermore, the generated assemblies in terms of contiguity and accuracy are on par with the other tools on most of the samples. Availability. HASLR is an open source tool available at https://github.com/vpc-ccg/haslr.</span></p><p>Address of the bookmark: <a href="https://github.com/vpc-ccg/haslr" rel="nofollow">https://github.com/vpc-ccg/haslr</a></p>]]></description>
	<dc:creator>BioStar</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/blog/view/43634/illumina-based-assembly-pipeline-steps</guid>
	<pubDate>Fri, 10 Dec 2021 06:22:54 -0600</pubDate>
	<link>https://bioinformaticsonline.com/blog/view/43634/illumina-based-assembly-pipeline-steps</link>
	<title><![CDATA[Illumina based assembly pipeline steps !]]></title>
	<description><![CDATA[<h3 id="illumina">Illumina<a href="https://nf-co.re/viralrecon#illumina"><span></span></a></h3><ol>
<li>Merge re-sequenced FastQ files (<a href="http://www.linfo.org/cat.html"><code>cat</code></a>)</li>
<li>Read QC (<a href="https://www.bioinformatics.babraham.ac.uk/projects/fastqc/"><code>FastQC</code></a>)</li>
<li>Adapter trimming (<a href="https://github.com/OpenGene/fastp"><code>fastp</code></a>)</li>
<li>Removal of host reads (<a href="http://ccb.jhu.edu/software/kraken2/"><code>Kraken 2</code></a>; <em>optional</em>)</li>
<li>Variant calling<ol>
<li>Read alignment (<a href="http://bowtie-bio.sourceforge.net/bowtie2/index.shtml"><code>Bowtie 2</code></a>)</li>
<li>Sort and index alignments (<a href="https://sourceforge.net/projects/samtools/files/samtools/"><code>SAMtools</code></a>)</li>
<li>Primer sequence removal (<a href="https://github.com/andersen-lab/ivar"><code>iVar</code></a>; <em>amplicon data only</em>)</li>
<li>Duplicate read marking (<a href="https://broadinstitute.github.io/picard/"><code>picard</code></a>; <em>optional</em>)</li>
<li>Alignment-level QC (<a href="https://broadinstitute.github.io/picard/"><code>picard</code></a>, <a href="https://sourceforge.net/projects/samtools/files/samtools/"><code>SAMtools</code></a>)</li>
<li>Genome-wide and amplicon coverage QC plots (<a href="https://github.com/brentp/mosdepth/"><code>mosdepth</code></a>)</li>
<li>Choice of multiple variant calling and consensus sequence generation routes (<a href="https://github.com/andersen-lab/ivar"><code>iVar variants and consensus</code></a>; <em>default for amplicon data</em> <em>||</em> <a href="http://samtools.github.io/bcftools/bcftools.html"><code>BCFTools</code></a>, <a href="https://github.com/arq5x/bedtools2/"><code>BEDTools</code></a>; <em>default for metagenomics data</em>)
<ul>
<li>Variant annotation (<a href="http://snpeff.sourceforge.net/SnpEff.html"><code>SnpEff</code></a>, <a href="http://snpeff.sourceforge.net/SnpSift.html"><code>SnpSift</code></a>)</li>
<li>Consensus assessment report (<a href="http://quast.sourceforge.net/quast"><code>QUAST</code></a>)</li>
<li>Lineage analysis (<a href="https://github.com/cov-lineages/pangolin"><code>Pangolin</code></a>)</li>
<li>Clade assignment, mutation calling and sequence quality checks (<a href="https://github.com/nextstrain/nextclade"><code>Nextclade</code></a>)</li>
<li>Individual variant screenshots with annotation tracks (<a href="https://asciigenome.readthedocs.io/en/latest/"><code>ASCIIGenome</code></a>)</li>
</ul>
</li>
<li>Intersect variants across callers (<a href="http://samtools.github.io/bcftools/bcftools.html"><code>BCFTools</code></a>)</li>
</ol></li>
<li><em>De novo</em> assembly<ol>
<li>Primer trimming (<a href="https://cutadapt.readthedocs.io/en/stable/guide.html"><code>Cutadapt</code></a>; <em>amplicon data only</em>)</li>
<li>Choice of multiple assembly tools (<a href="http://cab.spbu.ru/software/spades/"><code>SPAdes</code></a> <em>||</em> <a href="https://github.com/rrwick/Unicycler"><code>Unicycler</code></a> <em>||</em> <a href="https://github.com/GATB/minia"><code>minia</code></a>)
<ul>
<li>Blast to reference genome (<a href="https://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE_TYPE=BlastSearch"><code>blastn</code></a>)</li>
<li>Contiguate assembly (<a href="https://www.sanger.ac.uk/science/tools/pagit"><code>ABACAS</code></a>)</li>
<li>Assembly report (<a href="https://github.com/BU-ISCIII/plasmidID"><code>PlasmidID</code></a>)</li>
<li>Assembly assessment report (<a href="http://quast.sourceforge.net/quast"><code>QUAST</code></a>)</li>
</ul>
</li>
</ol></li>
<li>Present QC and visualisation for raw read, alignment, assembly and variant calling results (<a href="http://multiqc.info/"><code>MultiQC</code></a>)</li>
</ol>]]></description>
	<dc:creator>Surabhi Chaudhary</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/34618/mashmap-a-fast-and-approximate-software-for-mapping-long-reads-pacbioont-or-assembly-to-reference-genomes</guid>
	<pubDate>Tue, 12 Dec 2017 17:23:31 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/34618/mashmap-a-fast-and-approximate-software-for-mapping-long-reads-pacbioont-or-assembly-to-reference-genomes</link>
	<title><![CDATA[MashMap: a fast and approximate software for mapping long reads (PacBio/ONT) or assembly to reference genome(s)]]></title>
	<description><![CDATA[<p><span>MashMap is a fast and approximate software for mapping long reads (PacBio/ONT) or assembly to reference genome(s). It maps a query sequence against a reference region if and only if its estimated alignment identity is above a specified threshold. It does not compute the alignments explicitly, but rather estimates a&nbsp;</span><em>k</em><span>-mer based&nbsp;</span><a href="https://en.wikipedia.org/wiki/Jaccard_index">Jaccard similarity</a><span>&nbsp;using a combination of&nbsp;</span><a href="http://www.cs.princeton.edu/courses/archive/spr05/cos598E/bib/p76-schleimer.pdf">Winnowing</a><span>&nbsp;and&nbsp;</span><a href="https://en.wikipedia.org/wiki/MinHash">MinHash</a><span>. This is then converted to an estimate of sequence identity using the&nbsp;</span><a href="http://mash.readthedocs.org/">Mash</a><span>&nbsp;distance. An appropriate&nbsp;</span><em>k</em><span>-mer sampling rate is automatically determined given minimum local alignment length and identity thresholds. The efficiency of the algorithm improves as both of these thresholds are increased.</span></p><p>Address of the bookmark: <a href="https://github.com/marbl/MashMap" rel="nofollow">https://github.com/marbl/MashMap</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/40856/3d-de-novo-assembly-3d-dna-pipeline</guid>
	<pubDate>Sun, 02 Feb 2020 13:41:55 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/40856/3d-de-novo-assembly-3d-dna-pipeline</link>
	<title><![CDATA[3D de novo assembly (3D DNA) pipeline]]></title>
	<description><![CDATA[<p>For a detailed description of the pipeline and how it integrates with other tools designed by the Aiden Lab see&nbsp;<a href="http://aidenlab.org/assembly/manual_180322.pdf">Genome Assembly Cookbook</a>&nbsp;on&nbsp;<a href="http://aidenlab.org/assembly">http://aidenlab.org/assembly</a>.</p>
<p>For the original version of the pipeline and to reproduce the Hs2-HiC and the AaegL4 genomes reported in&nbsp;<a href="http://science.sciencemag.org/content/356/6333/92">(Dudchenko et al.,&nbsp;<em>Science</em>, 2017)</a>&nbsp;see the&nbsp;<a href="https://github.com/theaidenlab/3d-dna/tree/745779bdf64db6e55bddb70c24e9b58825938c33">original commit</a>.</p>
<p>For the detailed description of the merge section see&nbsp;<a href="https://github.com/theaidenlab/AGWG-merge">https://github.com/theaidenlab/AGWG-merge</a>.</p><p>Address of the bookmark: <a href="https://github.com/theaidenlab/3d-dna" rel="nofollow">https://github.com/theaidenlab/3d-dna</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/26306/busco</guid>
	<pubDate>Sun, 07 Feb 2016 16:02:39 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/26306/busco</link>
	<title><![CDATA[BUSCO]]></title>
	<description><![CDATA[<p>Assessing genome assembly and annotation completeness with Benchmarking Universal Single-Copy Orthologs</p>
<p>More at http://busco.ezlab.org/</p><p>Address of the bookmark: <a href="http://busco.ezlab.org/" rel="nofollow">http://busco.ezlab.org/</a></p>]]></description>
	<dc:creator>Jitendra Narayan</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/26906/paired-end-assembler-for-dna-sequences</guid>
	<pubDate>Wed, 06 Apr 2016 05:25:34 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/26906/paired-end-assembler-for-dna-sequences</link>
	<title><![CDATA[PAired-eND Assembler for DNA sequences]]></title>
	<description><![CDATA[<p>PANDASEQ is a program to align Illumina reads, optionally with PCR primers embedded in the sequence, and reconstruct an overlapping sequence.</p>
<p>&nbsp;</p>
<p>More at https://github.com/neufeld/pandaseq</p><p>Address of the bookmark: <a href="https://github.com/neufeld/pandaseq" rel="nofollow">https://github.com/neufeld/pandaseq</a></p>]]></description>
	<dc:creator>Neel</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/32730/ncbi-prokaryotic-genome-annotation-pipeline</guid>
	<pubDate>Tue, 16 May 2017 08:56:03 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/32730/ncbi-prokaryotic-genome-annotation-pipeline</link>
	<title><![CDATA[NCBI Prokaryotic Genome Annotation Pipeline]]></title>
	<description><![CDATA[<p>NCBI Prokaryotic Genome Annotation Pipeline is designed to annotate bacterial and archaeal genomes (chromosomes and plasmids).</p>
<p>Genome annotation is a multi-level process that includes prediction of protein-coding genes, as well as other functional genome units such as structural RNAs, tRNAs, small RNAs, pseudogenes, control regions, direct and inverted repeats, insertion sequences, transposons and other mobile elements.</p>
<p>NCBI has developed an automatic prokaryotic genome annotation pipeline that combines&nbsp;<em>ab initio</em>&nbsp;gene prediction algorithms with homology based methods. The first version of NCBI Prokaryotic Genome Automatic Annotation Pipeline (PGAAP;&nbsp;<a href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=pubmed&amp;dopt=Abstract&amp;list_uids=18416670">see Pubmed Article</a>) developed in 2005 has been replaced with an upgraded version that is capable of processing a larger data volume. You can find a more detailed description of the new version of&nbsp;the pipeline in&nbsp;<a href="https://www.ncbi.nlm.nih.gov/books/NBK174280/">NCBI Handbook chapter</a>. NCBI's annotation pipeline depends on several internal databases and is not currently available for download or use outside of the NCBI environment.</p>
<p>https://www.ncbi.nlm.nih.gov/genome/annotation_prok/</p><p>Address of the bookmark: <a href="https://www.ncbi.nlm.nih.gov/genome/annotation_prok/" rel="nofollow">https://www.ncbi.nlm.nih.gov/genome/annotation_prok/</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/40573/de-novo-genome-assembly-for-illumina-data</guid>
	<pubDate>Mon, 20 Jan 2020 05:13:29 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/40573/de-novo-genome-assembly-for-illumina-data</link>
	<title><![CDATA[De novo Genome Assembly for Illumina Data]]></title>
	<description><![CDATA[<p>Written and maintained by <a href="mailto:simon.gladman@unimelb.edu.au">Simon Gladman</a> - Melbourne Bioinformatics (formerly VLSCI)</p>
<p>Protocol Overview / Introduction</p>
<p>In this protocol we discuss and outline the process of de novo assembly for small to medium sized genomes.</p>
<p>https://www.melbournebioinformatics.org.au/tutorials/tutorials/assembly/assembly-protocol/</p><p>Address of the bookmark: <a href="https://www.melbournebioinformatics.org.au/tutorials/tutorials/assembly/assembly-protocol/" rel="nofollow">https://www.melbournebioinformatics.org.au/tutorials/tutorials/assembly/assembly-protocol/</a></p>]]></description>
	<dc:creator>Rahul Nayak</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/33847/omega2-metagenome-assembly-pipeline</guid>
	<pubDate>Mon, 10 Jul 2017 05:56:07 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/33847/omega2-metagenome-assembly-pipeline</link>
	<title><![CDATA[Omega2: metagenome assembly pipeline]]></title>
	<description><![CDATA[<p><span>Omega found overlaps between reads using a prefix/suffix hash table. The overlap graph of reads was simplified by removing transitive edges and trimming short branches. Unitigs were generated based on minimum cost flow analysis of the overlap graph and then merged to contigs and scaffolds using mate-pair information. In comparison with three de Bruijn graph assemblers (SOAPdenovo, IDBA-UD and MetaVelvet), Omega provided comparable overall performance on a HiSeq 100-bp dataset and superior performance on a MiSeq 300-bp dataset. In comparison with Celera on the MiSeq dataset, Omega provided more continuous assemblies overall using a fraction of the computing time of existing overlap-layout-consensus assemblers. This indicates Omega can more efficiently assemble longer Illumina reads, and at deeper coverage, for metagenomic datasets.</span></p><p>Address of the bookmark: <a href="http://omega.omicsbio.org/" rel="nofollow">http://omega.omicsbio.org/</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/pages/view/34418/spades-hybrid-genome-assembly</guid>
	<pubDate>Mon, 27 Nov 2017 08:05:40 -0600</pubDate>
	<link>https://bioinformaticsonline.com/pages/view/34418/spades-hybrid-genome-assembly</link>
	<title><![CDATA[SPAdes hybrid genome assembly]]></title>
	<description><![CDATA[<p>When you have both Illumina and Nanopore data, then SPAdes remains a good option for hybrid assembly - SPAdes was used to produce the&nbsp;<a href="https://gigascience.biomedcentral.com/articles/10.1186/s13742-015-0101-6">B fragilis assembly</a>&nbsp;by Mick Watson&rsquo;s group.</p><p>Again, running spades.py will show you the options:</p><div><pre><code>spades.py
</code></pre></div><p>This produces:</p><div><pre><code>SPAdes genome assembler v3.10.1

Usage: /usr/local/SPAdes-3.10.1-Linux/bin/spades.py [options] -o &lt;output_dir&gt;

Basic options:
-o      &lt;output_dir&gt;    directory to store all the resulting files (required)
--sc                    this flag is required for MDA (single-cell) data
--meta                  this flag is required for metagenomic sample data
--rna                   this flag is required for RNA-Seq data
--plasmid               runs plasmidSPAdes pipeline for plasmid detection
--iontorrent            this flag is required for IonTorrent data
--test                  runs SPAdes on toy dataset
-h/--help               prints this usage message
-v/--version            prints version

Input data:
--12    &lt;filename&gt;      file with interlaced forward and reverse paired-end reads
-1      &lt;filename&gt;      file with forward paired-end reads
-2      &lt;filename&gt;      file with reverse paired-end reads
-s      &lt;filename&gt;      file with unpaired reads
--pe&lt;#&gt;-12      &lt;filename&gt;      file with interlaced reads for paired-end library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--pe&lt;#&gt;-1       &lt;filename&gt;      file with forward reads for paired-end library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--pe&lt;#&gt;-2       &lt;filename&gt;      file with reverse reads for paired-end library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--pe&lt;#&gt;-s       &lt;filename&gt;      file with unpaired reads for paired-end library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--pe&lt;#&gt;-&lt;or&gt;    orientation of reads for paired-end library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9; &lt;or&gt; = fr, rf, ff)
--s&lt;#&gt;          &lt;filename&gt;      file with unpaired reads for single reads library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--mp&lt;#&gt;-12      &lt;filename&gt;      file with interlaced reads for mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--mp&lt;#&gt;-1       &lt;filename&gt;      file with forward reads for mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--mp&lt;#&gt;-2       &lt;filename&gt;      file with reverse reads for mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--mp&lt;#&gt;-s       &lt;filename&gt;      file with unpaired reads for mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--mp&lt;#&gt;-&lt;or&gt;    orientation of reads for mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9; &lt;or&gt; = fr, rf, ff)
--hqmp&lt;#&gt;-12    &lt;filename&gt;      file with interlaced reads for high-quality mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--hqmp&lt;#&gt;-1     &lt;filename&gt;      file with forward reads for high-quality mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--hqmp&lt;#&gt;-2     &lt;filename&gt;      file with reverse reads for high-quality mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--hqmp&lt;#&gt;-s     &lt;filename&gt;      file with unpaired reads for high-quality mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--hqmp&lt;#&gt;-&lt;or&gt;  orientation of reads for high-quality mate-pair library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9; &lt;or&gt; = fr, rf, ff)
--nxmate&lt;#&gt;-1   &lt;filename&gt;      file with forward reads for Lucigen NxMate library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--nxmate&lt;#&gt;-2   &lt;filename&gt;      file with reverse reads for Lucigen NxMate library number &lt;#&gt; (&lt;#&gt; = 1,2,..,9)
--sanger        &lt;filename&gt;      file with Sanger reads
--pacbio        &lt;filename&gt;      file with PacBio reads
--nanopore      &lt;filename&gt;      file with Nanopore reads
--tslr  &lt;filename&gt;      file with TSLR-contigs
--trusted-contigs       &lt;filename&gt;      file with trusted contigs
--untrusted-contigs     &lt;filename&gt;      file with untrusted contigs

Pipeline options:
--only-error-correction runs only read error correction (without assembling)
--only-assembler        runs only assembling (without read error correction)
--careful               tries to reduce number of mismatches and short indels
--continue              continue run from the last available check-point
--restart-from  &lt;cp&gt;    restart run with updated options and from the specified check-point ('ec', 'as', 'k&lt;int&gt;', 'mc')
--disable-gzip-output   forces error correction not to compress the corrected reads
--disable-rr            disables repeat resolution stage of assembling

Advanced options:
--dataset       &lt;filename&gt;      file with dataset description in YAML format
-t/--threads    &lt;int&gt;           number of threads
                                [default: 16]
-m/--memory     &lt;int&gt;           RAM limit for SPAdes in Gb (terminates if exceeded)
                                [default: 250]
--tmp-dir       &lt;dirname&gt;       directory for temporary files
                                [default: &lt;output_dir&gt;/tmp]
-k              &lt;int,int,...&gt;   comma-separated list of k-mer sizes (must be odd and
                                less than 128) [default: 'auto']
--cov-cutoff    &lt;float&gt;         coverage cutoff value (a positive float number, or 'auto', or 'off') [default: 'off']
--phred-offset  &lt;33 or 64&gt;      PHRED quality offset in the input reads (33 or 64)
                                [default: auto-detect]
</code></pre></div><p>As you can see this is also a &ldquo;pipeline&rdquo; of tools that can be switched on or off. SPAdes takes quite a long time, so for the purposes of this practical, something like this may suffice:</p><div><pre><code>spades.py -t 4 <span>\</span>
          -m 32 <span>\</span>
          -k 31,51,71 <span>\</span>
          --only-assembler <span>\</span>
          -1 miseq.1.fastq -2 miseq.2.fastq <span>\</span>
          --nanopore minion.fastq <span>\</span>
          -o hybrid_assembly
</code></pre></div><p>In turn, these parameters mean</p><ul>
<li>use 4 threads</li>
<li>max memory is 32Gb</li>
<li>use 3 kmer values to build the de bruijn graph(s) - 31, 51 and 71</li>
<li>only run the assembler, not the correction algorithm (for speed)</li>
<li>read 1 and read 2 of the MiSeq data</li>
<li>the nanopore data</li>
<li>put the output in folder &ldquo;hybrid_assembly&rdquo;</li>
</ul>]]></description>
	<dc:creator>Jit</dc:creator>
</item>

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