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	<title><![CDATA[BOL: Related items]]></title>
	<link>https://bioinformaticsonline.com/related/40994?offset=270</link>
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	<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/35055/jabba-hybrid-error-correction-for-long-sequencing-reads</guid>
	<pubDate>Fri, 05 Jan 2018 03:58:14 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/35055/jabba-hybrid-error-correction-for-long-sequencing-reads</link>
	<title><![CDATA[Jabba: Hybrid Error Correction for Long Sequencing Reads]]></title>
	<description><![CDATA[<p>Jabba is a hybrid error correction tool to correct third generation (PacBio / ONT) sequencing data, using second generation (Illumina) data.</p>
<p>Input</p>
<p>Jabba takes as input a concatenated de Bruijn graph and a set of sequences:</p>
<p>the de Bruijn graph should appear in fasta format with 1 entry per node, the meta information should be in the format:<br>&gt;NODE <br>the set of sequences should be in fasta or fastq format. These sequences will be corrected (e.g. PacBio reads). The corrections will be written to a file Jabba fasta.<br>The output is a file in fasta format with corrections of the long reads, and additionally a file in the input format containing uncorrected reads.</p>
<p>https://github.com/biointec/jabba/wiki</p>
<p>https://almob.biomedcentral.com/articles/10.1186/s13015-016-0075-7</p><p>Address of the bookmark: <a href="https://github.com/biointec/jabba" rel="nofollow">https://github.com/biointec/jabba</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/43828/understanding-hifi-reads</guid>
	<pubDate>Thu, 24 Mar 2022 19:48:11 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/43828/understanding-hifi-reads</link>
	<title><![CDATA[Understanding HiFi Reads !]]></title>
	<description><![CDATA[<p><span>While little public data is available for either of the new synthetic long read approaches, Illumina showed an example comparison earlier this year at the&nbsp;</span><a href="https://www.festivalofgenomics.com/rami-mehio" target="_blank">Festival of Genomics &amp; Biodata conference</a><span>&nbsp;(FoG 2022). In the IGV screenshot presented (below), synthetic Infinity reads &ndash; labeled &ldquo;Longas&rdquo; &ndash; are at the top, followed by standard Illumina short reads, and PacBio HiFi reads labeled &ldquo;CCS&rdquo; depicted at the bottom:</span></p><p>Address of the bookmark: <a href="http://pacb.com/blog/the-hifi-difference-true-long-reads-vs-synthetic-long-reads/" rel="nofollow">http://pacb.com/blog/the-hifi-difference-true-long-reads-vs-synthetic-long-reads/</a></p>]]></description>
	<dc:creator>Rahul Nayak</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/blog/view/44848/trust-but-verify-sequencing-your-cell-lines-might-reveal-an-uninvited-guest</guid>
	<pubDate>Wed, 04 Jun 2025 00:07:57 -0500</pubDate>
	<link>https://bioinformaticsonline.com/blog/view/44848/trust-but-verify-sequencing-your-cell-lines-might-reveal-an-uninvited-guest</link>
	<title><![CDATA[Trust But Verify: Sequencing Your Cell Lines Might Reveal an Uninvited Guest]]></title>
	<description><![CDATA[<p>High-throughput sequencing has become indispensable in cell biology, enabling detailed insights into chromatin structure, gene expression, and regulatory dynamics. Yet, when faced with unexpectedly low mapping rates to the human genome, researchers often rush to troubleshoot technical parameters&mdash;sequencer quality, adapter trimming, or aligner settings.</p><p>Before you go down that path, consider this critical biological question:<br /> <strong>Are you sequencing human cells&mdash;or bacterial contamination?</strong></p><h2>The Silent Saboteur: Mycoplasma in Cell Cultures</h2><p><em>Mycoplasma</em> contamination remains one of the most widespread and underdiagnosed issues in tissue culture work. Studies suggest that <strong>15&ndash;35% of cell lines in use may be contaminated</strong>, often without visible signs. Unlike other microbial infections, <em>Mycoplasma</em> does not produce cloudiness, odor, or a change in pH. Many researchers won&rsquo;t detect it unless they specifically test for it.</p><p>The consequences, however, are profound. <em>Mycoplasma</em> can significantly alter:</p><ul>
<li>
<p>Host gene expression patterns</p>
</li>
<li>
<p>Cell proliferation rates</p>
</li>
<li>
<p>Epigenetic profiles and chromatin accessibility</p>
</li>
<li>
<p>Cytokine signaling and immune responses</p>
</li>
</ul><p>In short, it can skew your results, compromise your biological conclusions, and invalidate weeks or months of research.</p><h2>A Simple Diagnostic Step: Map Against <em>Mycoplasma</em> Genomes</h2><p>If you encounter poor alignment rates to the human genome, consider mapping your reads to a <em>Mycoplasma</em> reference genome&mdash;or better yet, use a <strong>combined human + <em>Mycoplasma</em></strong> reference. There have been cases where over half of all reads, initially assumed to be from human cells, were in fact bacterial in origin. This check is fast, easy, and could save your project.</p><h2>How Contamination Happens&mdash;and Persists</h2><p><em>Mycoplasma</em> is small (0.1&ndash;0.3 &mu;m), lacks a cell wall, and can pass through standard filters undetected. Common sources include:</p><ul>
<li>
<p>Contaminated reagents (e.g., FBS)</p>
</li>
<li>
<p>Infected cell lines obtained from other labs</p>
</li>
<li>
<p>Poor aseptic technique or shared equipment</p>
</li>
</ul><p>Once present, it spreads quickly between cultures and can persist for months, silently affecting results.</p><h2>Why Treatment Is Difficult</h2><p>While antibiotics such as Plasmocin or BM-Cyclin are sometimes used, they often offer only partial resolution and may themselves alter cell behavior. In many cases, the best course of action is to <strong>discard the contaminated culture</strong> and start with a fresh, verified stock.</p><h2>Practical Recommendations for Researchers</h2><ul>
<li>
<p><strong>Routinely test for <em>Mycoplasma</em></strong> using PCR, qPCR, or fluorescence-based assays</p>
</li>
<li>
<p><strong>Incorporate contamination screens into your sequencing QC pipeline</strong></p>
</li>
<li>
<p><strong>Use combined reference genomes</strong> when mapping ambiguous reads</p>
</li>
<li>
<p><strong>Practice strict aseptic technique</strong> and monitor all incoming cell lines</p>
</li>
<li>
<p><strong>Don&rsquo;t ignore unexplained data anomalies</strong>&mdash;they might point to contamination</p>
</li>
</ul><h2>Closing Thought: Contamination Is a Biological Variable</h2><p>It&rsquo;s easy to view poor mapping as a technical issue, but sometimes the problem lies deeper&mdash;in the biology itself. <em>Mycoplasma</em> contamination doesn&rsquo;t just interfere with sequencing; it interferes with science. As a research community, we must treat contamination not as an afterthought, but as a key variable to control.</p><p>So next time your reads won&rsquo;t align, don&rsquo;t just tune the aligner. Ask if your cells are telling the truth&mdash;or if they're hiding something.</p>]]></description>
	<dc:creator>LEGE</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/2726/comparison-of-short-read-de-novo-alignment-algorithms</guid>
	<pubDate>Wed, 21 Aug 2013 07:56:01 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/2726/comparison-of-short-read-de-novo-alignment-algorithms</link>
	<title><![CDATA[Comparison of Short Read De Novo Alignment Algorithms]]></title>
	<description><![CDATA[<p>Excellent article to introduce different sequencing methods along with tools for de novo assembly of sequencing reads and their relevant references.</p>
<p>Title:&nbsp;<strong>Comparison of Short Read De Novo Alignment Algorithms&nbsp;</strong></p>
<p>Author<strong>: Nikhil Gopal</strong></p><p>Address of the bookmark: <a href="http://biochem218.stanford.edu/Projects%202011/Gopal%202011.pdf" rel="nofollow">http://biochem218.stanford.edu/Projects%202011/Gopal%202011.pdf</a></p>]]></description>
	<dc:creator>Rahul Agarwal</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/36865/perga-a-paired-end-read-guided-de-novo-assembler-for-extending-contigs-using-svm-and-look-ahead-approach</guid>
	<pubDate>Tue, 05 Jun 2018 09:57:11 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/36865/perga-a-paired-end-read-guided-de-novo-assembler-for-extending-contigs-using-svm-and-look-ahead-approach</link>
	<title><![CDATA[PERGA: A Paired-End Read Guided De Novo Assembler for Extending Contigs Using SVM and Look Ahead Approach]]></title>
	<description><![CDATA[PERGA - Paired End Reads Guided Assembler

PERGA is a novel sequence reads guided de novo assembly approach which adopts greedy-like prediction strategy for assembling reads to contigs and scaffolds. Instead of using single-end reads to construct contig, PERGA uses paired-end reads and different read overlap sizes from O ≥ Omax to Omin to resolve the gaps and branches. Moreover, by constructing a decision model using machine learning approach based on branch features, PERGA can determine the correct extension in 99.7% of cases. PERGA will try to extend the contigs by all feasible nucleotides and determine if these multiple extensions due to sequencing errors or repeats by using looking ahead technology, and it also try to separate the different repeats of nearby genomic regions to make the assembly result more longer and accurate.

The simulated E.coli paired-end reads data are generated using GemSim (KE McElroy, F Luciani, T Thomas. Gemsim: General, Error-Model Based Simulator of Next-Generation Sequencing Data. BMC Genomics 2012, 13:74), with coverage 50x, 60x, 100x, read lengths 100-bp, and can be downloaded from https://github.com/zhuxiao/data_PERGA.<p>Address of the bookmark: <a href="https://github.com/hitbio/PERGA" rel="nofollow">https://github.com/hitbio/PERGA</a></p>]]></description>
	<dc:creator>Rahul Nayak</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/37221/asplice-a-scalable-and-memory-efficient-algorithm-for-de-novo-transcriptome-assembly</guid>
	<pubDate>Tue, 03 Jul 2018 04:09:46 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/37221/asplice-a-scalable-and-memory-efficient-algorithm-for-de-novo-transcriptome-assembly</link>
	<title><![CDATA[ASplice: a scalable and memory-efficient algorithm for de novo transcriptome assembly]]></title>
	<description><![CDATA[With increased availability of de novo assembly algorithms, it is feasible to study entire transcriptomes of non-model organisms. While algorithms are available that are specifically designed for performing transcriptome assembly from high-throughput sequencing data, they are very memory-intensive, limiting their applications to small data sets with few libraries.

Texas A&amp;M University researchers develop a transcriptome assembly algorithm that recovers alternatively spliced isoforms and expression levels while utilizing as many RNA-Seq libraries as possible that contain hundreds of gigabases of data. New techniques are developed so that computations can be performed on a computing cluster with moderate amount of physical memory.

Availability – A software program that implements the algorithm is available at: http://faculty.cse.tamu.edu/shsze/asplice.

Sze SH, Pimsler ML, Tomberlin JK, Jones CD, Tarone AM. (2017) A scalable and memory-efficient algorithm for de novo transcriptome assembly of non-model organisms. BMC Genomics 18(Suppl 4):387.<p>Address of the bookmark: <a href="http://faculty.cse.tamu.edu/shsze/asplice/" rel="nofollow">http://faculty.cse.tamu.edu/shsze/asplice/</a></p>]]></description>
	<dc:creator>Rahul Nayak</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/38212/megahit-an-ultra-fast-single-node-solution-for-large-and-complex-metagenomics-assembly-via-succinct-de-bruijn-graph</guid>
	<pubDate>Wed, 14 Nov 2018 04:50:27 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/38212/megahit-an-ultra-fast-single-node-solution-for-large-and-complex-metagenomics-assembly-via-succinct-de-bruijn-graph</link>
	<title><![CDATA[MEGAHIT: an ultra-fast single-node solution for large and complex metagenomics assembly via succinct de Bruijn graph]]></title>
	<description><![CDATA[<p><span>MEGAHIT is a single node assembler for large and complex metagenomics NGS reads, such as soil. It makes use of succinct&nbsp;</span><em>de Bruijn</em><span>&nbsp;graph (SdBG) to achieve low memory assembly. MEGAHIT can&nbsp;</span><span>optionally</span><span>&nbsp;utilize a CUDA-enabled GPU to accelerate its SdBG contstruction. The GPU-accelerated version of MEGAHIT has been tested on NVIDIA GTX680 (4G memory) and Tesla K40c (12G memory) with CUDA 5.5, 6.0 and 6.5. MEGAHIT v1.0 or greater also supports IBM Power PC and has been tested on IBM POWER8.</span></p>
<p><span>https://academic.oup.com/bioinformatics/article/31/10/1674/177884</span></p><p>Address of the bookmark: <a href="https://github.com/voutcn/megahit" rel="nofollow">https://github.com/voutcn/megahit</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/39098/sda-long-read-sequence-and-assembly-of-segmental-duplications</guid>
	<pubDate>Tue, 05 Mar 2019 10:00:57 -0600</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/39098/sda-long-read-sequence-and-assembly-of-segmental-duplications</link>
	<title><![CDATA[SDA: Long-read sequence and assembly of segmental duplications]]></title>
	<description><![CDATA[<p><span><span>Segmental Duplication Assembler (SDA; https://github.com/mvollger/SDA) constructs graphs in which paralogous sequence variants define the nodes and long-read sequences provide attraction and repulsion edges, enabling the partition and assembly of long reads corresponding to distinct paralogs.<br></span></span></p>
<p><span><span>https://github.com/mvollger/SDA</span></span></p><p>Address of the bookmark: <a href="https://www.nature.com/articles/s41592-018-0236-3" rel="nofollow">https://www.nature.com/articles/s41592-018-0236-3</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>
<item>
	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/40099/contiguator</guid>
	<pubDate>Fri, 04 Oct 2019 01:27:58 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/40099/contiguator</link>
	<title><![CDATA[CONTIGuator !]]></title>
	<description><![CDATA[<p><span>CONTIGuator is a Python script for Linux environments whose purpose is to speed-up the bacterial genome assembly process and to obtain a first insight of the genome structure using the well-known artemis comparison tool (ACT).</span></p>
<p>&nbsp;</p><p>Address of the bookmark: <a href="https://sourceforge.net/projects/contiguator/" rel="nofollow">https://sourceforge.net/projects/contiguator/</a></p>]]></description>
	<dc:creator>BioStar</dc:creator>
</item>
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	<guid isPermaLink="true">https://bioinformaticsonline.com/bookmarks/view/41734/supernova-generates-phased-whole-genome-de-novo-assemblies-from-a-chromium-prepared-library</guid>
	<pubDate>Sun, 31 May 2020 01:59:30 -0500</pubDate>
	<link>https://bioinformaticsonline.com/bookmarks/view/41734/supernova-generates-phased-whole-genome-de-novo-assemblies-from-a-chromium-prepared-library</link>
	<title><![CDATA[Supernova: generates phased, whole-genome de novo assemblies from a Chromium-prepared library.]]></title>
	<description><![CDATA[<p>Supernova generates phased, whole-genome&nbsp;<em>de novo</em>&nbsp;assemblies from a Chromium-prepared library.</p>
<p>Please see&nbsp;<a href="https://support.10xgenomics.com/de-novo-assembly/guidance/doc/achieving-success-with-de-novo-assembly">Achieving Success with De Novo Assembly</a>&nbsp;and&nbsp;<a href="https://support.10xgenomics.com/de-novo-assembly/software/overview/system-requirements">System Requirements</a>&nbsp;<em>before</em>&nbsp;creating your Chromium libraries for assembly.</p>
<p>Supernova should be run using 38-56x coverage of the genome.<br>&bull; Somewhat higher coverage is&nbsp;<em>sometimes</em>&nbsp;advantageous.<br>&bull; Supernova will exit if it finds that coverage is far from the recommended range.<br>&bull; Note that at most 2.14 billion reads are allowed.<br>&bull; Please note that we have not extensively tested genomes larger than human, and any genome above approximately 4 GB should be considered experimental and is not supported.</p><p>Address of the bookmark: <a href="https://support.10xgenomics.com/de-novo-assembly/software/pipelines/latest/using/running" rel="nofollow">https://support.10xgenomics.com/de-novo-assembly/software/pipelines/latest/using/running</a></p>]]></description>
	<dc:creator>Jit</dc:creator>
</item>

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