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	<title><![CDATA[BOL: Related items]]></title>
	<link>https://bioinformaticsonline.com/related/37502?offset=320</link>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/blog/view/42633/protocol-for-de-novo-genome-assembly-using-illumina-reads</guid>
	<pubDate>Sat, 16 Jan 2021 21:42:11 -0600</pubDate>
	<link>https://bioinformaticsonline.com/blog/view/42633/protocol-for-de-novo-genome-assembly-using-illumina-reads</link>
	<title><![CDATA[Protocol for De novo Genome Assembly using Illumina Reads]]></title>
	<description><![CDATA[<p>In this protocol, we address and describe the de novo assembly method for small to medium-sized genomes.</p><p><strong>What is de novo genome assembly?<br /></strong>The method of taking a large number of short DNA sequences and placing them back together to create a reflection of the original chromosomes from which the DNA originated relates to genome assembly. No previous knowledge of the source DNA sequence length, structure or composition is inferred by De novo genome assemblies. The DNA of the target organism is split up into millions of tiny parts and read on a sequencing computer in a genome sequencing experiment. Depending on the sequencing system used, these "reads" range from 20 to 1000 nucleotide base pairs (bp) in length. Usually, length reads of 36 - 150 bp are produced for Illumina style short read sequencing. These reads can be either &ldquo;single ended&rdquo; as described above or &ldquo;paired end.&rdquo;</p><p><strong>Why genome assembly?</strong><br />In basic research into why and how they live, as well as in applied topics, identifying the DNA sequence of an organism is useful. Awareness of a DNA sequence may be useful in virtually any biological research because of the relevance of DNA to living things. For example, it may be used in medicine to classify, diagnose and eventually improve genetic disorder therapies. Similarly, pathogens study can lead to treatments for infectious diseases.</p><p><strong>Raw NGS data</strong><br />Reads can be saved as a Fasta file as text or in a FastQ file with their attributes.&nbsp;FastQ is the most common read file format since this is what the Illumina sequencing pipeline creates. This will henceforth be the subject of our conversation.</p><p><strong>In a nutshell the protocol:</strong> <br />Get the sequence file(s) read from the sequencing machine (s). <br />Look at the readings - have an idea of what you have and what the standard is like. <br />If required, raw data cleanup/quality trimming. <br />Choose an adequate parameter set for assembly. <br />Assemble the data into scaffolds/contigs. <br />Examine the assembly performance and determine the efficiency of the assembly.</p><p><strong>Read Quality Control:</strong><br />Check the qualiy with fastQC.<br />Script<br />https://bioinformaticsonline.com/snippets/view/42540/install-fastqc-using-conda</p><p>Quality trimming/cleanup of read files.<br />This function trims adapters, barcodes and other contaminants from the reads.<br />Script<br />https://bioinformaticsonline.com/snippets/view/42542/trimmomatic-command</p><p><strong>Genome Assembly:</strong><br />The object of this portion of the protocol is to explain the method of assembling the reads trimmed by quality into draft contigs.</p><blockquote><p>spades.py -1 illumina_R1.fastq.gz -2 illumina_R2.fastq.gz --careful --cov-cutoff auto -o result_of_spades_assembly_all_illumina</p></blockquote><p>A significant range of short-read assemblers are available. Everyone with strengths and disadvantages of their own. <br /><em>Some of the assemblers available include:</em><br />Velvet<br />SOAP-denovo<br />MIRA<br />ALLPATHS</p><p>Next step is to assess the suitability and what to do with a draft package of contiguous details for the remainder of the study now.&nbsp;Few stuff you can note about the contigs you just created:&nbsp;They're the draft Contigs. Any mis-assemblies can occur.</p><p><strong>Mis-assembly checking and assembly metric tools:</strong><br />QUAST - Quality assessment tool for genome assembly http://bioinf.spbau.ru/quast<br />Mauve assembly metrics - http://code.google.com/p/ngopt/wiki/How_To_Score_Genome_Assemblies_with_Mauve<br />InGAP-SV - https://sites.google.com/site/nextgengenomics/ingap and http://ingap.sourceforge.net/<br />inGAP is also useful for finding structural variants between genomes from read mappings.</p><p><strong>Genome finishing tools:</strong><br />Semi-automated gap fillers:<br />Gap filler - http://www.baseclear.com/landingpages/basetools-a-wide-range-of-bioinformatics-solutions/gapfiller/</p><p>IMAGE (V2) - http://sourceforge.net/apps/mediawiki/image2/index.php?title=Main_Page</p><p><strong>Genome visualisers and editors:</strong><br />Artemis - http://www.sanger.ac.uk/resources/software/artemis/<br />IGV - http://www.broadinstitute.org/igv/</p><p><strong>Automated and semi automated annotation tools:</strong><br />Prokka - https://github.com/tseemann/prokka<br />RAST - http://www.nmpdr.org/FIG/wiki/view.cgi/FIG/RapidAnnotationServer<br />JCVI Annotation Service - http://www.jcvi.org/cms/research/projects/annotation-service/</p><p><strong>Frequent command use for the analysis are at:</strong></p><p>https://bioinformaticsonline.com/blog/view/38765/list-of-tools-frequently-used-while-genome-assembly<br />https://bioinformaticsonline.com/pages/view/42275/frequent-parameters-for-bioinformatics-tools</p>]]></description>
	<dc:creator>BioStar</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/news/view/14191/scalpel</guid>
	<pubDate>Wed, 20 Aug 2014 02:07:58 -0500</pubDate>
	<link>https://bioinformaticsonline.com/news/view/14191/scalpel</link>
	<title><![CDATA[Scalpel]]></title>
	<description><![CDATA[<p>A team from Cold Spring Harbor Laboratory has released an algorithm, called Scalpel, for finding insertions and deletions in next generation sequencing data sets. Scalpel, which is open source and <a href="http://scalpel.sourceforge.net/" title="available for download">available for download</a> on SourceForge,&nbsp;<span>outperformed the popular tools GATK HaplotypeCaller and SOAPindel in test runs on both simulated and real whole human exomes.</span></p><p>Like other indel callers, Scalpel works by performing <em>de novo</em>&nbsp;assembly of regions of interest, so that misalignment to the reference genome cannot obscure the presence of an insertion or deletion. Scalpel's innovation is to repeatedly check its assembly before comparing to the reference genome, to account for simple sequence repeats that are a regular source of error in indel calling. When Scalpel assembles an exon, it collects reads that map to that exon (including partial matches), splits them into k-mers, and creates a de Bruijn graph to span the exon; however, if it detects repeats in the map, it iteratively increases the size of the k-mers by one base until the repeats are eliminated. This ensures that the final assembly of the exon is highly accurate while minimizing compute time.</p><p>The Cold Spring Harbor team's validation of Scalpel, <a href="http://www.nature.com/nmeth/journal/vaop/ncurrent/full/nmeth.3069.html" title="published over the weekend in Nature Methods">published over the weekend in <em>Nature Methods</em></a>, compares Scalpel's performance on a live whole exome against HaplotypeCaller and SOAPindel. The donor is an individual with serious neurological disorders, which may be linked to a high incidence of indels. One thousand indels from this individual's exome, called by one or more of the informatics pipelines, were selected for focused resequencing. This resequencing revealed a 77% true positive rate for Scalpel calls, dramatically better than the rates for either of the competing tools; Scalpel performed especially well with indels longer than five base pairs, a traditional weak point for indel callers.</p><p>Finally, the authors demonstrate Scalpel's use on a large set of genetic data from nearly 600 families who donated samples to the Simons Simplex Collection, a project of the Simons Foundation Autism Research Initiative. Scalpel found a very high enrichment for indels in children affected by autism, compared with their unaffected siblings, a pattern that persisted even after excluding common variants.</p>]]></description>
	<dc:creator>Shruti Paniwala</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/news/view/27344/orffinder-with-smart-blast</guid>
	<pubDate>Tue, 17 May 2016 01:43:15 -0500</pubDate>
	<link>https://bioinformaticsonline.com/news/view/27344/orffinder-with-smart-blast</link>
	<title><![CDATA[ORFfinder with smart BLAST]]></title>
	<description><![CDATA[<p><span>ORF Finder</span></p><p><span><a href="http://www.ncbi.nlm.nih.gov/orffinder">ORFfinder</a><span>&nbsp;is a graphical analysis tool for finding open reading frames (ORFs). We&rsquo;ve been working on a few updates, and we&rsquo;d like to find out what you think about them. Read on to find out what you can do with the new ORFfinder.</span></span></p><p>Smart BLAST (https://ncbiinsights.ncbi.nlm.nih.gov/2015/07/29/smartblast/)</p><p>Select one or a group of ORFs and BLAST several databases at once, and use the newly developed&nbsp;<a href="http://blast.ncbi.nlm.nih.gov/smartblast/">SmartBLAST</a>&nbsp;to verify protein names.&nbsp;Looking for the traditional results from&nbsp;<a href="http://blast.ncbi.nlm.nih.gov/Blast.cgi">BLAST</a>? They&rsquo;re there too.</p>]]></description>
	<dc:creator>Jit</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/30680/easybuild</guid>
	<pubDate>Fri, 27 Jan 2017 16:00:43 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/30680/easybuild</link>
	<title><![CDATA[EasyBuild]]></title>
	<description><![CDATA[<p><a href="https://github.com/hpcugent/easybuild">EasyBuild</a><span>&nbsp;is a software build and installation framework that allows you to manage (scientific) software on High Performance Computing (HPC) systems in an efficient way.</span><br><span>A full list of supported software packages is available&nbsp;</span><a href="http://easybuild.readthedocs.io/en/latest/version-specific/Supported_software.html">here</a><span>.</span></p><p>Address of the bookmark: <a href="https://hpcugent.github.io/easybuild/" rel="nofollow">https://hpcugent.github.io/easybuild/</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/34678/svfinder-tool-for-detecting-genomic-rearrangement-form-dna-seq-data</guid>
	<pubDate>Thu, 14 Dec 2017 15:51:40 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/34678/svfinder-tool-for-detecting-genomic-rearrangement-form-dna-seq-data</link>
	<title><![CDATA[SVfinder: Tool for detecting genomic rearrangement form DNA-seq data]]></title>
	<description><![CDATA[<p><span>SVfinder provides genome-wide detection of structural variants from next generation paired-end sequencing reads.</span></p><p>Address of the bookmark: <a href="https://github.com/cauyrd/SVfinder" rel="nofollow">https://github.com/cauyrd/SVfinder</a></p>]]></description>
	<dc:creator>Robert M Willioms</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/36012/gmol-an-interactive-tool-for-3d-genome-structure-visualization</guid>
	<pubDate>Wed, 21 Mar 2018 12:25:20 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/36012/gmol-an-interactive-tool-for-3d-genome-structure-visualization</link>
	<title><![CDATA[GMOL: An Interactive Tool for 3D Genome Structure Visualization]]></title>
	<description><![CDATA[<p><span>GMOL was developed based upon our multi-scale approach that allows a user to scale between six separate levels within the genome. With GMOL, a user can choose any unit at any scale and scale it up or down to visualize its structure and retrieve corresponding genome sequences.</span></p><p>Address of the bookmark: <a href="https://www.nature.com/articles/srep20802" rel="nofollow">https://www.nature.com/articles/srep20802</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/42204/g-nest-the-gene-neighborhood-scoring-tool</guid>
	<pubDate>Fri, 25 Sep 2020 20:09:18 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/42204/g-nest-the-gene-neighborhood-scoring-tool</link>
	<title><![CDATA[G-NEST: The Gene NEighborhood Scoring Tool]]></title>
	<description><![CDATA[<p><span>The Gene NEighborhood Scoring Tool (G-NEST) combines genomic location, gene expression, and evolutionary sequence conservation data to score putative gene neighborhoods across all window sizes. Primary author of final code = William F. Martin. Example data files are in the separate repository.</span></p><p>Address of the bookmark: <a href="https://github.com/dglemay/G-NEST" rel="nofollow">https://github.com/dglemay/G-NEST</a></p>]]></description>
	<dc:creator>Neel</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/44659/figeno-tool-for-plotting-sequencing-data-along-genomic-coordinates</guid>
	<pubDate>Tue, 17 Sep 2024 02:28:15 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/44659/figeno-tool-for-plotting-sequencing-data-along-genomic-coordinates</link>
	<title><![CDATA[Figeno: Tool for plotting sequencing data along genomic coordinates.]]></title>
	<description><![CDATA[<p><span>Tool for plotting sequencing data along genomic coordinates.</span></p>
<div>
<pre><code>FIGENO is a
  FIGure
    GENerator
for GENOmics</code></pre>
</div>
<p dir="auto">With figeno, you can plot various types of sequencing data along genomic coordinates. Video overview:&nbsp;<a href="https://www.youtube.com/watch?v=h1cBeXoSYTA">https://www.youtube.com/watch?v=h1cBeXoSYTA</a>.</p>
<p dir="auto"><a href="https://github.com/CompEpigen/figeno/blob/main/docs/content/images/figeno.png" target="_blank"><img src="https://github.com/CompEpigen/figeno/raw/main/docs/content/images/figeno.png" alt="figeno" style="border: 0px;"></a></p><p>Address of the bookmark: <a href="https://github.com/CompEpigen/figeno" rel="nofollow">https://github.com/CompEpigen/figeno</a></p>]]></description>
	<dc:creator>LEGE</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/35420/telomerehunter</guid>
	<pubDate>Fri, 02 Feb 2018 04:23:59 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/35420/telomerehunter</link>
	<title><![CDATA[TelomereHunter]]></title>
	<description><![CDATA[<p><span>TelomereHunter is a tool for estimating telomere content from human whole-genome sequencing data. It is designed to take BAM files from a tumor and a matching control sample as input. However, it is also possible to run TelomereHunter with one input file. TelomereHunter extracts and sorts telomeric reads from the input sample(s). For the estimation of telomere content, GC biases are taken into account. Finally, the results of TelomereHunter are visualized in several diagrams.</span><br><br><span>TelomereHunter is available for download at the following address:&nbsp;</span><a href="https://pypi.python.org/pypi/telomerehunter/" target="_blank">https://pypi.python.org/pypi/telomerehunter/</a></p><p>Address of the bookmark: <a href="http://www.dkfz.de/en/applied-bioinformatics/telomerehunter/telomerehunter.html" rel="nofollow">http://www.dkfz.de/en/applied-bioinformatics/telomerehunter/telomerehunter.html</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/34380/chsminer-a-gui-tool-to-identify-chromosomal-homologous-segments</guid>
	<pubDate>Sat, 18 Nov 2017 16:55:49 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/34380/chsminer-a-gui-tool-to-identify-chromosomal-homologous-segments</link>
	<title><![CDATA[CHSMiner: a GUI tool to identify chromosomal homologous segments]]></title>
	<description><![CDATA[<div id="ASec1">
<h3>Background</h3>
<p>The identification of chromosomal homologous segments (CHS) within and between genomes is essential for comparative genomics. Various processes including insertion/deletion and inversion could cause the degeneration of CHSs.</p>
</div>
<div id="ASec2">
<h3>Results</h3>
<p>Here we present a Java software CHSMiner that detects CHSs based on shared gene content alone. It implements fast greedy search algorithm and rigorous statistical validation, and its friendly graphical interface allows interactive visualization of the results. We tested the software on both simulated and biological realistic data and compared its performance with similar existing software and data source.</p>
</div>
<div id="ASec3">
<h3>Conclusion</h3>
<p>CHSMiner is characterized by its integrated workflow, fast speed and convenient usage. It will be useful for both experimentalists and bioinformaticians interested in the structure and evolution of genomes.</p>
<p>&nbsp;</p>
<p>https://github.com/zhenwang100/CHSMiner</p>
</div><p>Address of the bookmark: <a href="https://almob.biomedcentral.com/articles/10.1186/1748-7188-4-2" rel="nofollow">https://almob.biomedcentral.com/articles/10.1186/1748-7188-4-2</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
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