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	<title><![CDATA[BOL: Related items]]></title>
	<link>https://bioinformaticsonline.com/related/44722?offset=240</link>
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	<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/blog/view/44227/common-methods-to-discover-tandem-repeats</guid>
	<pubDate>Thu, 09 Mar 2023 02:40:52 -0600</pubDate>
	<link>https://bioinformaticsonline.com/blog/view/44227/common-methods-to-discover-tandem-repeats</link>
	<title><![CDATA[Common methods to discover tandem repeats]]></title>
	<description><![CDATA[<div><div><div><div><div><div><div><div><div><div><p>Tandem repeats are DNA sequences that are repeated in a contiguous manner in the genome. These sequences are often used as genetic markers and are important in many areas of genetics and genomics research. Here are some methods for discovering tandem repeats in genomes:</p><ol>
<li>
<p>Tandem Repeat Finder: Tandem Repeat Finder is a software tool that identifies tandem repeats in DNA sequences. It is available for free download and can be used on both nucleotide and protein sequences. The tool uses a statistical algorithm to identify repeats based on their length, copy number, and overall composition.</p>
</li>
<li>
<p>RepeatMasker: RepeatMasker is another software tool that can identify tandem repeats in DNA sequences. It works by comparing the input sequence to a database of known repeats and then identifies any tandem repeats that match those in the database.</p>
</li>
<li>
<p>PCR-based methods: Polymerase chain reaction (PCR) can be used to amplify and detect tandem repeats in genomic DNA. PCR primers are designed to flank the tandem repeat region, and amplification of the target DNA fragment can be visualized on a gel. This method can be useful for detecting novel tandem repeats and for genotyping.</p>
</li>
<li>
<p>Southern blotting: Southern blotting is a classic method for detecting DNA fragments in a sample. It can be used to detect tandem repeats by digesting genomic DNA with a restriction enzyme, separating the fragments by gel electrophoresis, and then probing the blot with a tandem repeat-specific probe.</p>
</li>
</ol><p>Overall, a combination of these methods can be used to comprehensively identify tandem repeats in genomes.</p></div></div></div></div></div></div></div></div></div></div>]]></description>
	<dc:creator>BioStar</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/pages/view/27321/slurm-basics</guid>
	<pubDate>Fri, 13 May 2016 04:42:24 -0500</pubDate>
	<link>https://bioinformaticsonline.com/pages/view/27321/slurm-basics</link>
	<title><![CDATA[SLURM basics !]]></title>
	<description><![CDATA[<p><a href="http://bioinformaticsonline.com/bookmarks/view/27238/slurm" target="_blank">SLURM</a> is a queue management system and stands for Simple Linux Utility for Resource Management. SLURM was developed at the Lawrence Livermore National Lab and currently runs some of the largest compute clusters in the world.</p><p>SLURM is similar in many ways to most other queue systems. You write a batch script then submit it to the queue manager. The queue manager then schedules your job to run on the queue (or partition in SLURM parlance) that you designate. Below we will provide an outline of how to submit jobs to SLURM, how SLURM decides when to schedule your job and how to monitor progress.</p><p>SLURM has a number of valuable features compared to other job management systems:</p><ul>
<li><em>Kill and Requeue</em> SLURM&rsquo;s ability to kill and requeue is superior to that of other systems. It waits for jobs to be cleared before scheduling the high priority job. It also does kill and requeue on memory rather than just on core count.</li>
<li><em>Memory</em> Memory requests are sacrosanct in SLURM. Thus the amount of memory you request at run time is guaranteed to be there. No one can infringe on that memory space and you cannot exceed the amount of memory that you request.</li>
<li><em>Accounting Tools</em> SLURM has a back end database which stores historical information about the cluster. This information can be queried by the users who are curious about how much resources they have used.</li>
</ul><p><strong>Summary of SLURM commands</strong></p><p>The table below shows a summary of SLURM commands. These commands are described in more detail below along with links to the SLURM doc site.</p><table>
<tbody>
<tr><th>&nbsp;</th><th>SLURM</th><th>SLURM Example</th></tr>
<tr>
<td>Submit a batch serial job</td>
<td><a href="http://slurm.schedmd.com/sbatch.html">sbatch</a></td>
<td><code>sbatch runscript.sh</code></td>
</tr>
<tr>
<td>Run a script interatively</td>
<td><a href="http://slurm.schedmd.com/srun.html">srun</a></td>
<td><code>srun --pty -p interact -t 10 --mem 1000 /bin/bash /bin/hostname</code></td>
</tr>
<tr>
<td>Kill a job</td>
<td><a href="http://slurm.schedmd.com/scancel.html">scancel</a></td>
<td><code>scancel 999999</code></td>
</tr>
<tr>
<td>View status of queues</td>
<td><a href="http://slurm.schedmd.com/squeue.html">squeue</a></td>
<td><code>squeue -u akitzmiller</code></td>
</tr>
<tr>
<td>Check current job by id</td>
<td><a href="http://slurm.schedmd.com/squeue.html">sacct</a></td>
<td><code>sacct -j 999999</code></td>
</tr>
</tbody>
</table>]]></description>
	<dc:creator>Radha Agarkar</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/blog/view/44720/a-beginners-guide-to-using-kraken-for-taxonomic-classification</guid>
	<pubDate>Fri, 13 Dec 2024 11:29:03 -0600</pubDate>
	<link>https://bioinformaticsonline.com/blog/view/44720/a-beginners-guide-to-using-kraken-for-taxonomic-classification</link>
	<title><![CDATA[A Beginner&#039;s Guide to Using Kraken for Taxonomic Classification]]></title>
	<description><![CDATA[<div>Kraken is a popular bioinformatics tool designed for fast and accurate taxonomic classification of metagenomic sequences. Its efficiency and precision make it a go-to resource for analyzing microbial communities, including bacteria, viruses, archaea, and fungi. Whether you're new to bioinformatics or experienced in the field, Kraken is an indispensable tool for taxonomic analysis.</div><div><div><div><div dir="auto"><div><div><p>In this blog, we&rsquo;ll walk through the basics of Kraken, from installation to running an analysis, and highlight its key features and applications.</p><h4><strong>What is Kraken?</strong></h4><p>Kraken is a sequence classification tool that assigns taxonomic labels to DNA sequences using exact k-mer matching. It uses a reference database of genomes, dividing sequences into k-mers and identifying matches in a computationally efficient way.</p><h4><strong>Key Features of Kraken</strong></h4><ul>
<li><strong>Speed</strong>: Kraken processes data much faster than alignment-based methods.</li>
<li><strong>Accuracy</strong>: It uses a precise k-mer matching algorithm for high-resolution taxonomic assignments.</li>
<li><strong>Scalability</strong>: It can handle large metagenomic datasets.</li>
<li><strong>Custom Databases</strong>: You can build and use custom databases tailored to your research needs.</li>
</ul><h4><strong>Installing Kraken</strong></h4><ol>
<li>
<p><strong>System Requirements</strong></p>
<ul>
<li>A Unix-based operating system (Linux/macOS).</li>
<li>Sufficient computational resources for database building (RAM and disk space).</li>
</ul>
</li>
<li>
<p><strong>Installation Steps</strong></p>
<ul>
<li>Clone the Kraken repository from GitHub:
<div>
<div>&nbsp;</div>
<div dir="ltr"><code>git <span style="font-size: 12.8px; font-weight: normal;">clone</span> https://github.com/DerrickWood/kraken.git <span style="font-size: 12.8px; font-weight: normal;">cd</span> kraken </code></div>
</div>
</li>
<li>Compile the Kraken binaries:
<div>
<div>&nbsp;</div>
<div dir="ltr"><code>make </code></div>
</div>
</li>
<li>Add Kraken to your PATH for easy access:
<div>
<div>&nbsp;</div>
<div dir="ltr"><code><span style="font-size: 12.8px; font-weight: normal;">export</span> PATH=<span style="font-size: 12.8px; font-weight: normal;">$PATH</span>:/path/to/kraken </code></div>
</div>
</li>
</ul>
</li>
</ol><h4><strong>Preparing a Database</strong></h4><p>Kraken requires a database of reference genomes. You can use a pre-built database or create a custom one.</p><ol>
<li>
<p><strong>Downloading a Pre-built Database</strong><br />Kraken offers pre-built databases, such as the <em>MiniKraken</em> database, which is lightweight and suitable for smaller datasets. Download it using:</p>
<div>
<div dir="ltr"><code>kraken-build --download-library minikraken </code></div>
</div>
</li>
<li>
<p><strong>Building a Custom Database</strong><br />To include specific genomes, download FASTA files and build the database:</p>
<div>
<div dir="ltr"><code>kraken-build --download-library bacteria --threads 4 --db my_database kraken-build --build --db my_database </code></div>
</div>
<p>This process may take considerable time and resources, depending on the size of the database.</p>
</li>
</ol><h4><strong>Running Kraken</strong></h4><p>Once the database is ready, you can classify sequences.</p><ol>
<li>
<p><strong>Basic Usage</strong><br />Use the following command to classify sequences:</p>
<div>
<div dir="ltr"><code>kraken --db my_database --threads 4 --fastq-input input_sequences.fastq --output kraken_output.txt </code></div>
</div>
<p>Key options:</p>
<ul>
<li><code>--db</code>: Specifies the database.</li>
<li><code>--threads</code>: Number of threads for parallel processing.</li>
<li><code>--fastq-input</code>: Indicates input file format (FASTQ/FASTA).</li>
</ul>
</li>
<li>
<p><strong>Interpreting Results</strong><br />Kraken generates an output file with columns for sequence IDs, taxonomic classifications, and the confidence score.</p>
</li>
</ol><h4><strong>Visualizing Kraken Results</strong></h4><p>Kraken results can be visualized using tools like <strong>Krona</strong> or converted to human-readable reports using <code>kraken-report</code>.</p><ol>
<li>
<p><strong>Generate a Report</strong></p>
<div>
<div dir="ltr"><code>kraken-report --db my_database kraken_output.txt &gt; kraken_report.txt </code></div>
</div>
</li>
<li>
<p><strong>Krona Visualization</strong><br />Install Krona and convert Kraken output for visualization:</p>
<div>
<div dir="ltr"><code>cut -f2,3 kraken_output.txt | ktImportTaxonomy -o krona_output.html </code></div>
</div>
<p>Open the HTML file in your browser to interactively explore the taxonomic classifications.</p>
</li>
</ol><h4><strong>Advanced Usage</strong></h4><ol>
<li>
<p><strong>Confidence Thresholds</strong><br />Adjust the confidence threshold for classification using the <code>--confidence</code> option. Higher values reduce false positives but may miss some true positives:</p>
<div>
<div dir="ltr"><code>kraken --db my_database --confidence 0.1 --fastq-input input.fastq </code></div>
</div>
</li>
<li>
<p><strong>Paired-End Reads</strong><br />For paired-end sequencing data, use:</p>
<div>
<div dir="ltr"><code>kraken --db my_database --paired reads_1.fastq reads_2.fastq </code></div>
</div>
</li>
<li>
<p><strong>Customizing K-mers</strong><br />Kraken allows you to set custom k-mer lengths during database building for specific applications.</p>
</li>
</ol><h4><strong>Applications of Kraken</strong></h4><ul>
<li><strong>Microbial Ecology</strong>: Characterizing microbial communities in soil, water, and the human microbiome.</li>
<li><strong>Pathogen Detection</strong>: Identifying pathogens in clinical samples.</li>
<li><strong>Fungal Research</strong>: Analyzing fungal diversity in metagenomic datasets.</li>
<li><strong>Environmental Monitoring</strong>: Tracking microbial populations in diverse habitats.</li>
</ul><h4><strong>Conclusion</strong></h4><p>Kraken is a versatile and efficient tool for taxonomic classification in metagenomics. Its speed, accuracy, and flexibility make it a favorite among bioinformaticians. By following this guide, you can set up and use Kraken to unlock insights into microbial and fungal communities, paving the way for discoveries in ecology, medicine, and biotechnology.</p></div></div></div></div></div></div>]]></description>
	<dc:creator>Neel</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/pages/view/36711/ancestral-sequence-reconstruction-steps</guid>
	<pubDate>Fri, 18 May 2018 08:28:26 -0500</pubDate>
	<link>https://bioinformaticsonline.com/pages/view/36711/ancestral-sequence-reconstruction-steps</link>
	<title><![CDATA[Ancestral sequence reconstruction steps !]]></title>
	<description><![CDATA[<div><strong>Ancestral sequence reconstruction</strong>&nbsp;(<strong>ASR</strong>) &ndash; also known as&nbsp;<strong>ancestral gene</strong>/<strong>sequence reconstruction</strong>/<strong>resurrection</strong>&nbsp;&ndash; is a technique used in the study of&nbsp;molecular evolution. The method consists of the synthesis of an ancestral&nbsp;gene&nbsp;and expression of the corresponding ancestral&nbsp;protein.&nbsp;<a href="https://en.wikipedia.org/wiki/Ancestral_sequence_reconstruction#cite_note-thornton-1"></a>The idea of protein 'resurrection' was suggested in 1963 by Pauling and Zuckerkandl.<a href="https://en.wikipedia.org/wiki/Ancestral_sequence_reconstruction#cite_note-2"></a>&nbsp;Some early efforts were made in the eighties-nineties, led by the laboratory of&nbsp;Steven A. Benner, showing the potential of this technique &ndash; one that only started to be fulfilled in the post-genomic era.<a href="https://en.wikipedia.org/wiki/Ancestral_sequence_reconstruction#cite_note-3"></a>&nbsp;Thanks to the improvement of algorithms and of better sequencing and synthesis techniques, the method was developed further in the early 2000s to allow the resurrection of a greater variety of and much more ancient genes.<a href="https://en.wikipedia.org/wiki/Ancestral_sequence_reconstruction#cite_note-4"></a>&nbsp;Over the last decade, ancestral protein resurrection has developed as a strategy to reveal the mechanisms and dynamics of protein evolution.&nbsp;</div><div>&nbsp;</div><div>BEAST is the best way to predict the ancestral structure. but, I suggest following steps?</div><div>&nbsp;</div><div>1- Alignments "Mafft -&nbsp;<a href="https://www.researchgate.net/deref/http%3A%2F%2Fmafft.cbrc.jp%2Falignment%2Fsoftware%2Fsource.html" target="_blank">http://mafft.cbrc.jp/alignment/software/source.html</a>"</div><div>mafft --maxiterate 1000 --reorder --thread 24 --genafpair Dataset.fasta &gt; Dataset_Alig.fasta</div><div>&nbsp;</div><div>2- Your dataset has a good phylogenetic signal, is possible to perform with Tree-Puzzle "<a href="https://www.researchgate.net/deref/http%3A%2F%2Fwww.tree-puzzle.de" target="_blank">http://www.tree-puzzle.de</a>";</div><div>&nbsp;</div><div id="yui_3_14_1_1_1526649596608_1443">3 - This dataset which the saturation index, I perform with "<a href="https://www.researchgate.net/deref/http%3A%2F%2Fdambe.bio.uottawa.ca%2Fdambe.asp" target="_blank">http://dambe.bio.uottawa.ca/dambe.asp</a>";</div><div>&nbsp;</div><div>4- Has evidence of possible recombination in your dataset, the evaluate if this presence or absence, because this may to influence the grouping of clades, I perform with</div><div>---recombination</div><div>&nbsp;</div><div>4.1- Phi-test, implemented in SplitTree4"<a href="https://www.researchgate.net/deref/http%3A%2F%2Fwww.splitstree.org" target="_blank">http://www.splitstree.org</a>", (.nex file)</div><div>&nbsp;</div><div>4.2- GARD deployed in webserver in the DataMonkey "<a href="https://www.researchgate.net/deref/http%3A%2F%2Fwww.datamonkey.org%2F" target="_blank">http://www.datamonkey.org/</a>" - turning to the amino acid seaview -&gt; view proteins -&gt; save as ...) Ideally do a tree-based groups.</div><div>&nbsp;</div><div>4.3- RDP4 for download and installation on Windows in "<a href="https://www.researchgate.net/deref/http%3A%2F%2Fweb.cbio.uct.ac.za%2F~darren%2Frdp.html" target="_blank">http://web.cbio.uct.ac.za/~darren/rdp.html</a>"</div><div>&nbsp;</div><div>4.4- Hyphy (Mac, Windows, Linux) in "<a href="https://www.researchgate.net/deref/http%3A%2F%2Fhyphy.org%2Fw%2Findex.php%2FDownload" target="_blank">http://hyphy.org/w/index.php/Download</a>"</div><div>&nbsp;</div><div>4.5- Path-o-Gen (temporal structure of a tree input file -&gt; arquivo.tre)</div><div>These steps above, I call of pre-processing to inferences phylogenetic...</div><div>&nbsp;</div><div>5- Perform phylogenetic tree, used Bayesian Inference with Molecular Clock, but is necessary Clock Testing:</div><div>&nbsp;</div><div>- This step is performed with program Beast (Beauti, Beast and TreeAnnotator), and Tracer_v1.5 more FigTree to inspection.</div><div>&nbsp;</div><div>- Tutorials:&nbsp;<a href="https://www.researchgate.net/deref/http%3A%2F%2Fbeast.bio.ed.ac.uk%2Ftutorials" target="_blank">http://beast.bio.ed.ac.uk/tutorials</a></div><div>- Downloads:&nbsp;<a href="https://www.researchgate.net/deref/http%3A%2F%2Fbeast.bio.ed.ac.uk%2Fdownloads" target="_blank">http://beast.bio.ed.ac.uk/downloads</a></div>]]></description>
	<dc:creator>Surabhi Chaudhary</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/bookmarks/view/34416/miniasm-very-fast-olc-based-de-novo-assembler-for-noisy-long-reads</guid>
	<pubDate>Mon, 27 Nov 2017 07:58:49 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/34416/miniasm-very-fast-olc-based-de-novo-assembler-for-noisy-long-reads</link>
	<title><![CDATA[miniasm: very fast OLC-based de novo assembler for noisy long reads]]></title>
	<description><![CDATA[<p>Miniasm is a very fast OLC-based&nbsp;<em>de novo</em>&nbsp;assembler for noisy long reads. It takes all-vs-all read self-mappings (typically by&nbsp;<a href="https://github.com/lh3/minimap">minimap</a>) as input and outputs an assembly graph in the&nbsp;<a href="https://github.com/pmelsted/GFA-spec/blob/master/GFA-spec.md">GFA</a>&nbsp;format. Different from mainstream assemblers, miniasm does not have a consensus step. It simply concatenates pieces of read sequences to generate the final&nbsp;<a href="http://wgs-assembler.sourceforge.net/wiki/index.php/Celera_Assembler_Terminology">unitig</a>&nbsp;sequences. Thus the per-base error rate is similar to the raw input reads.</p>
<p>So far miniasm is in early development stage. It has only been tested on a dozen of PacBio and Oxford Nanopore (ONT) bacterial data sets. Including the mapping step, it takes about 3 minutes to assemble a bacterial genome. Under the default setting, miniasm assembles 9 out of 12 PacBio datasets and 3 out of 4 ONT datasets into a single contig. The 12 PacBio data sets are&nbsp;<a href="https://github.com/PacificBiosciences/DevNet/wiki/E.-coli-Bacterial-Assembly">PacBio E. coli sample</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS473430">ERS473430</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS544009">ERS544009</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS554120">ERS554120</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS605484">ERS605484</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS617393">ERS617393</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS646601">ERS646601</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS659581">ERS659581</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS670327">ERS670327</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS685285">ERS685285</a>,&nbsp;<a href="http://www.ebi.ac.uk/ena/data/view/ERS743109">ERS743109</a>&nbsp;and a&nbsp;<a href="https://github.com/PacificBiosciences/DevNet/wiki/E.-coli-20kb-Size-Selected-Library-with-P6-C4/ce0533c1d2a957488594f0b29da61ffa3e4627e8">deprecated PacBio E. coli data set</a>. ONT data are acquired from the&nbsp;<a href="http://lab.loman.net/2015/09/24/first-sqk-map-006-experiment/">Loman Lab</a>.</p>
<p>For a&nbsp;<em>C. elegans</em>&nbsp;<a href="https://github.com/PacificBiosciences/DevNet/wiki/C.-elegans-data-set">PacBio data set</a>&nbsp;(only 40X are used, not the whole dataset), miniasm finishes the assembly, including reads overlapping, in ~10 minutes with 16 CPUs. The total assembly size is 105Mb; the N50 is 1.94Mb. In comparison, the&nbsp;<a href="https://github.com/PacificBiosciences/Bioinformatics-Training/wiki/HGAP">HGAP3</a>produces a 104Mb assembly with N50 1.61Mb.&nbsp;<a href="http://lh3lh3.users.sourceforge.net/download/ce-miniasm.png">This dotter plot</a>&nbsp;gives a global view of the miniasm assembly (on the X axis) and the HGAP3 assembly (on Y). They are broadly comparable. Of course, the HGAP3 consensus sequences are much more accurate. In addition, on the whole data set (assembled in ~30 min), the miniasm N50 is reduced to 1.79Mb. Miniasm still needs improvements.</p>
<p>Miniasm confirms that at least for high-coverage bacterial genomes, it is possible to generate long contigs from raw PacBio or ONT reads without error correction. It also shows that&nbsp;<a href="https://github.com/lh3/minimap">minimap</a>&nbsp;can be used as a read overlapper, even though it is probably not as sensitive as the more sophisticated overlapers such as&nbsp;<a href="https://github.com/marbl/MHAP">MHAP</a>&nbsp;and&nbsp;<a href="https://github.com/thegenemyers/DALIGNER">DALIGNER</a>. Coupled with long-read error correctors and consensus tools, miniasm may also be useful to produce high-quality assemblies.</p>
<p>Minimap and miniasm are ultrafast tools for (i) mapping and (ii) assembly. Designed for long, noisy reads, they do not have a correction or consensus step, and therefore the resulting assemblies are contiguous (i.e. long) but very noisy (i.e. full of errors)</p>
<p>We start with an all against all comparison:</p>
<div>
<pre><code>minimap -Sw5 -L100 -m0 -t8 reads.fq reads.fq | gzip -1 &gt; reads.paf.gz
</code></pre>
</div>
<p>Then we can assemble</p>
<div>
<pre><code>miniasm -f reads.fq reads.paf.gz &gt; reads.gfa
</code></pre>
</div>
<p>Convert GFA to FASTA:</p>
<div>
<pre><code>awk <span>'/^S/{print "&gt;"$2"\n"$3}'</span> reads.gfa | fold &gt; reads.fa
</code></pre>
</div>
<p>And then count how many contigs:</p>
<div>
<pre><code>grep <span>"&gt;"</span> reads.fa | wc -l</code></pre>
</div>
<p>&nbsp;</p>
<pre><span><span>#</span> Download sample PacBio from the PBcR website</span>
wget -O- http://www.cbcb.umd.edu/software/PBcR/data/selfSampleData.tar.gz <span>|</span> tar zxf -
ln -s selfSampleData/pacbio_filtered.fastq reads.fq
<span><span>#</span> Install minimap and miniasm (requiring gcc and zlib)</span>
git clone https://github.com/lh3/minimap <span>&amp;&amp;</span> (cd minimap <span>&amp;&amp;</span> make)
git clone https://github.com/lh3/miniasm <span>&amp;&amp;</span> (cd miniasm <span>&amp;&amp;</span> make)
<span><span>#</span> Overlap</span>
minimap/minimap -Sw5 -L100 -m0 -t8 reads.fq reads.fq <span>|</span> gzip -1 <span>&gt;</span> reads.paf.gz
<span><span>#</span> Layout</span>
miniasm/miniasm -f reads.fq reads.paf.gz <span>&gt;</span> reads.gfa</pre><p>Address of the bookmark: <a href="https://github.com/lh3/miniasm" rel="nofollow">https://github.com/lh3/miniasm</a></p>]]></description>
	<dc:creator>Jit</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/35345/rgfa-powerful-and-convenient-handling-of-assembly-graphs</guid>
	<pubDate>Thu, 25 Jan 2018 05:47:53 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/35345/rgfa-powerful-and-convenient-handling-of-assembly-graphs</link>
	<title><![CDATA[RGFA: powerful and convenient handling of assembly graphs]]></title>
	<description><![CDATA[<p><span>RGFA, an implementation of the proposed GFA specification in Ruby. It allows the user to conveniently parse, edit and write GFA files. Complex operations such as the separation of the implicit instances of repeats and the merging of linear paths can be performed. A typical application of RGFA is the editing of a graph, to finish the assembly of a sequence, using information not available to the assembler. We illustrate a use case, in which the assembly of a repetitive metagenomic fosmid insert was completed using a script based on RGFA.</span></p>
<p><span>https://github.com/ggonnella/rgfa</span></p><p>Address of the bookmark: <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5103826/" rel="nofollow">https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5103826/</a></p>]]></description>
	<dc:creator>Rahul Nayak</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/36867/cerulean-a-hybrid-assembly-using-high-throughput-short-and-long-reads</guid>
	<pubDate>Tue, 05 Jun 2018 10:10:15 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/36867/cerulean-a-hybrid-assembly-using-high-throughput-short-and-long-reads</link>
	<title><![CDATA[Cerulean: A hybrid assembly using high throughput short and long reads]]></title>
	<description><![CDATA[Cerulean extends contigs assembled using short read datasets like Illumina paired-end reads using long reads like PacBio RS long reads.

Cerulean v0.1 has been implemented with bacterial genomes in mind.

The method is fully described in Deshpande, V., Fung, E. D., Pham, S., &amp; Bafna, V. (2013). Cerulean: A hybrid assembly using high throughput short and long reads. arXiv preprint arXiv:1307.7933.
http://arxiv.org/abs/1307.7933<p>Address of the bookmark: <a href="https://sourceforge.net/projects/ceruleanassembler/" rel="nofollow">https://sourceforge.net/projects/ceruleanassembler/</a></p>]]></description>
	<dc:creator>Rahul Nayak</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/37554/finishersca-repeat-aware-tool-for-upgrading-de-novo-assembly-using-long-reads</guid>
	<pubDate>Mon, 20 Aug 2018 04:08:50 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/37554/finishersca-repeat-aware-tool-for-upgrading-de-novo-assembly-using-long-reads</link>
	<title><![CDATA[FinisherSC:a repeat-aware tool for upgrading de novo assembly using long reads]]></title>
	<description><![CDATA[<p><br>Here is the command to run the tool:</p>
<pre><code>python finisherSC.py destinedFolder mummerPath
</code></pre>
<p>If you are running on server computer and would like to use multiple threads, then the following commands can generate 20 threads to run FinisherSC.</p>
<pre><code>python finisherSC.py -par 20 destinedFolder mummerPath
</code></pre>
<p>Sometimes, if the names of raw reads and contigs consists of special characters/formats, FinisherSC/MUMmer may not parse them correctly. In that case, you want to have a quick renaming of the names of contigs/reads in contigs.fasta or raw_reads.fasta using the following command.</p>
<pre><code>    perl -pe 's/&gt;[^\$]*$/"&gt;Seg" . ++$n ."\n"/ge' raw_reads.fasta &gt; newRaw_reads.fasta
    cp newRaw_reads.fasta raw_reads.fasta
    perl -pe 's/&gt;[^\$]*$/"&gt;Seg" . ++$n ."\n"/ge' contigs.fasta &gt; newContigs.fasta
    cp newContigs.fasta contigs.fasta</code></pre><p>Address of the bookmark: <a href="https://github.com/kakitone/finishingTool" rel="nofollow">https://github.com/kakitone/finishingTool</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>

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