Questions tagged [genome-sequencing]

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Interpreting 'samtools mpileup' output for multiple inputs

I would like to calculate sequencing coverage for a WGS project. Both long and short reads. I've used samtools as following: ...
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Locate the saved template of minknow

I am using minknow on ubuntu. After saving the template (settings), where is that file locally stored in the system? Also what is its format/extension?
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2 votes
2 answers
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Is this a complete bacterial genome? (CheckM)

I've been working with a bacterial genome assembly. The initial stats look pretty good (3 main pieces, depth ~50x [20x - 100x] if I calculated it right). I'd expect this to be a virtually complete ...
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Pros and cons between genomes and transcriptomes?

What are the major differences between the use cases of a shot-gun genome and an mRNA capture transcriptome? Especially when it comes to downstream analyses such as looking for orthology, and ...
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Length of Contigs in Transcriptome and Whole Genome Assembly

Why are there shorter contigs from transcriptome assembly than from a whole genome assembly? I know the difference between transcriptome and genome, but don't really understand what contigs are in the ...
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Can someone help me estimating the runtime of the pipeline applied by the vertebrate genome project?

The vertebrate genome project (VGP) has a lot of interesting publications such as this one. The rough pipeline is outlined below: Here the pipeline in more detail: While the paper describes all the ...
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Modeling number of reads mapped to a gene

I am looking for a probability distribution of a number of reads mapped to a particular gene in metagenomic sequencing (NGS, shotgun, likely illumina). Naively one could model it via a binomial (or ...
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1 vote
1 answer
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Which one is a more convenient assembly?

I have developed a software for de novo genome assembly. Its performance varies gradually according to how much data you employ. At initial stages it often produces contigs that look like that when ...
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2 votes
2 answers
236 views

Genome assembly of SRR12196449 with SPAdes

I am trying to assemble the run SRR12196449 with SPAdes. The description of their project is: This project expected to standardize a method for amplification and ...
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2 votes
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Assembly reads having a copy of their beginning in their tail

I am analyzing the reads for the SARS-CoV-2 assembly with id SRR11140748. Apparently these reads were obtained with parallel sequencing by Illumina and Oxford Nanopore Technologies. I have found these ...
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3 votes
1 answer
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Overlap in Paired-end Reads for Sequencing?

Regarding Sequencing: Do/can the paired-end reads have overlaps?
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1 vote
1 answer
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Location of Reads in Sequencing

I have questions regarding sequencing: Are paired-end reads from different stands? What about single reads, are they coming from both strands?
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Warning in fastqc

I am checking the quality control of my sequences using the Fastqc tool. For some steps, like "Per base GC content", etc., I received a warning. So, I was wondering whether I Should also take care of ...
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How do researchers account for heterozygosity in genome assembly?

this is a very general question about diploid genome assembly. I am wondering how people deal with heterozygous region in a genome when assembling ? Do they pick one of the haplotype to be in the ...
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Extract reads from bam files by their @RG

How could I extract reads from bam files by their read groups @RG ?, I've got one file containing all reads for 5 samples, the SM is appears NONE, So, I want to extract each sample reads by the @RG ...
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3 votes
2 answers
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Produce a single sequential FASTA sequence out of BAM

I'm having problems properly looking for a solution because I'm a layman in Bioinformatics not familiar with the terminology. I'm hoping you can nudge me in the right direction, please! Thank you very ...
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3 votes
2 answers
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Separation of mixed plasmid DNA sequences post whole-plasmid sequencing

Imagine a DNA sample containing a mixture of different intact plasmids. These samples are sequenced using either MiSeq or HiSeq sequencing. Would it possible to assemble these plasmids post-sequencing ...
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1 answer
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What are 2D reads in the Oxford MinIon?

Reading the Mash: fast genome and metagenome distance estimation paper, I see the reference to "2d reads" notion under the minION context (a sequencing technique) e.g ...
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1 answer
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Generating basic dna sequences in R

Looking for some guidance. In an introductory bioinformatics class I am tasked with doing the following in R: Create the sequence ACTGACTG Generate a random string of nucleotides with equal ...
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Genotyping with Illumina HumanOmni1-Quad before whole genome sequencing

I read a paper about (whole genome sequencing analysis of 100 Southeast Asian Malays. It says "Prior to library preparation, each sample was genotyped on the Illumina HumanOmni1-Quad as an initial ...
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Can I run STAR without an annotation file?

I wish to use Rascaf to scaffold a fragmented draft genome. For this, I need to provide a BAM file of aligned RNA-seq reads and the draft genome. So, I indexed the draft genome with STAR like this: <...
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1 vote
1 answer
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How to QC overamplified shotgun library?

I made some NEB ultra II whole-genome shotgun libraries with around ~200ng of template going into the indexing PCR step. The PCR was run for 13 cycles and I ended up having high library concentrations ...
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9 votes
4 answers
6k views

What are the pros and cons of the different basecallers in Oxford Nanopore Technology Sequencing?

What are the pros and cons of the different basecallers in Oxford Nanopore Technology Sequencing? I am about to start a MinION run on my laptop. What should I consider when choosing my basecaller? ...
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3 votes
1 answer
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Can the canu assembler output a fastq file of the final assembly just like HGAP4?

I have assembled some genome from Sequel PacBio data both with HGAP4 on the SMRT Link interface and using canu on the command line. The HGAP4 assembler outputs a fastq file of the final assembly such ...
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9 votes
2 answers
5k views

How do PCR duplicates arise and why is it important to remove them for NGS analysis?

I am trying to understand PCR duplicates in NGS analyses (actually whole-genome). I searched, and the best answer I found is in this blog. However I don't understand if I understood how PCR ...
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15 votes
3 answers
937 views

Publicly available genome sequence database for viruses?

As a small introductory project, I want to compare genome sequences of different strains of influenza virus. What are the publicly available databases of influenza virus gene/genome sequences?
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15 votes
2 answers
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Downloading a reference Genome for Bowtie2

How do I download a reference genome that I can use with bowtie2? Specifically HG19. On UCSC there are a lot of file options.
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24 votes
4 answers
754 views

Tools for simulating Oxford Nanopore reads

Are there any free open source software tools available for simulating Oxford Nanopore reads?
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