Questions tagged [alignment]
These questions are about sequence alignment.
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Why does Nucmer Genome alignment produce faulty Dot plot?
I am using Mummer v.3.23 to make a dot plot between two genomes of the same species. One genome was assembled using PacBio and one from Dovetail. None of them is ...
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1answer
34 views
bowtie No alignments
This question was also asked on Biostars
hi all, I am working on miRNA alignment using bowtie. But it seems to fail to map. Here are the commands:
...
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4answers
61 views
Remove gaps from alignment?
I have an MSA (protein sequence) and I have used various programmes (Clustal, Aliview, MEGA11 etc) to align. However, in all programmes I get many gaps which is not ideal as I am trying to construct a ...
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1answer
52 views
Why does BWA MEM orientation contradict my library prep method
I have some RNA-seq data from a stranded paired end library prep, with dUTP and UDG preparation, so the orientation should be RF (confirmed with sequencing provider). I assembled the reads with ...
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13 views
Global vs Local alignment scoring matrix
When creating scoring matrices for global and local alignments is there are difference regarding the "highest score of the matrix". For example, for the two matrices below, the correct ...
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45 views
How to convert the chromosome position for insertions/deletions to rsIDs in a GWAS summary statistics?
I am trying to convert chromosome positions to rsIDs of a GWAS summary statistics file. I used bedtools intersect to merge the reference genome GRCh37 and the ...
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18 views
Type and number of gaps in sequence alignments
I used Kalign and Muscle to align given sequences, the results are presented below. My question is, based on the two alignments which tool seems better? I would personally believe that the Muscle tool ...
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1answer
28 views
blastn returning an inferior alignment
In my work I've found that in circumstances where there is a mismatch near the edges of the query sequence, blastn prefers to return a shorter contiguous alignment, rather than allowing for a mismatch ...
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1answer
17 views
How do you search for new enzymes that are more stable for handling, immobilization?
Noob here. I get that I should deduce what characteristics the ideal new enzyme should meet, and then use tools such as PDB and blast to compare to the old enzymes, and use other tools such as pymol, ...
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26 views
Alignment with inserts and keeping the indexing of ref seq intact
Parts of sequences are given below-
Reference sequence (pre-alignment):
ATTAAAGGTTTATACCTTCCCAGGTAACAAACCAACCAACTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGT
...
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25 views
use Kallisto in galaxy
I want to use kalisto for sequence alignment in Galaxy. i found this field empty: there is no option available for the reference transcriptome.
how I can get one?
Can I let the other configuration by ...
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1answer
18 views
Total pairs of amino acid substitution
I am reading a blog and it says:
The numbers for identities and replacements used for calculating the overall alignment score in the expression above are usually presented in the form of a 20 x 20 ...
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1answer
55 views
where to find sample contig data
Where or How can I find contigs? I am trying to learn Bioinformatics and I want to make a reference-based gene search. I also want to align contigs with a given reference sequence. From NCBI other ...
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1answer
43 views
File format of substitution matrix in clustalw
I need to set the substitution matrix used by command line CLUSTALW when comparing DNA sequences to:
0 -1 -1 -1
-1 0 -1 -1
-1 -1 0 -1
-1 -1 -1 0
from my ...
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1answer
31 views
Understanding ViennaRNA RNAdistance scoring table
I'm trying to compare the output of 2 different algorithms of RNA structure prediction
(my implementation of Nussinov vs RNA-mfold algorithm) using the RNAdistance algorithm that is part of ViennaRNA ...
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2answers
130 views
Calculate genome coverage and depth from alignment
I have a .bam alignment file and a genome reference .fasta file. I am looking for a easy to use tool (that I can reference in a publication) to calculate the percentage coverage of the reference by ...
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1answer
177 views
STAR aligner multiple fastq files
I’m using STAR to align fastq files from SMART-seq2. I have raw data folder containing sub-folders with samples names the sub-folders each contain fastq file. How can I make a bash command in order ...
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1answer
54 views
Median string problem & multiple sequence alignment
I read about the median string problem as an introduction to the multiple sequence alignment, however none of the MSA algorithms used seems to be using the idea of finding the median string.
To my ...
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1answer
84 views
Global alignment between two sequence X and Y with maximum number of identical matches
If 2 protein sequence X of length m and Y of length n and if there are several highest scoring global alignments, then I want to get one alignment that has largest number of columns in which a letter ...
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1answer
67 views
Supplementary aligments in VAF
This question has also been asked on Biostars
I have a doubt, are supplementary alignment usually considered when the variant allele frequency is calculated? Thanks a lot.
In my case I have some ...
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1answer
136 views
Assessing PyMol sequence alignment object
I use cealign in PyMol for structure alignment. Instead of visualization, I want to return the alignment object to my python script for further analysis. Is there a function to return the object ...
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1answer
28 views
Measure the purifying selection for certain taxa along a phylogeny
I am posting this message because I need clarity about the analysis I want to perform.
In my analysis, I have a homologous gene in 13 species, and I would like to evaluate the selection pressure of ...
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2answers
47 views
What programs account for structural alignment of different parts of distant homologs which have significant structural differences?
If there is a need to perform structural alignment of different parts of distant homologs, which program one should use?
Since distant homologs often have significant structural changes, meaning the ...
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1answer
136 views
TopHat2 versus HISAT2 inner workings
In my intro to bioinformatics course, we mentioned that TopHat2 and HISAT2 will both try to align as many reads as possible to the reference genome (TopHat2 has been superseded by HISAT2). For the ...
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31 views
BAM file filteing to remain best isoform
I ran HiSat2, MarkDuplicate, removed reads with the lower quality score than 40 and finally only kept properly paired reads.
After the BAM filtering steps, I used the Scallop results with TransDecoder....
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1answer
30 views
Installation of PRANK MSA in WLS Ubuntu 20.04 LTS
I want to install PRANK on the Windows 10 Linux subsystem (Ubuntu 20.04 LTS), I have followed the installation instructions to no avail.
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1answer
117 views
why in RNA seq don't we only use reference transcriptome?
I would like to ask why in RNA seq analysis (alignment step) we use sometimes reference genome instead of reference transcriptome? thank you!
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What are the symbols of $*$ and '$\_$' in an unknown alignment format for HLA data from the IMGT dataset?
Has anyone worked with the IMGT HLA database/dataset before? IMGT-HLA git repo they have some convenient text files (eg. link to gene A .txt file) with the genomic data for the different alleles of ...
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1answer
315 views
Difference between genome assembly and genome sequence alignment to a reference to find structural variants
I'm trying to determine what the difference and benefits of genome assembly and genome sequence alignments are when trying to identify structural variants or transposons in populations.
I've been ...
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1answer
44 views
Display a score or graph to visualize the degree of conservation of each residue from alignment data
I would like to display a score or graph to visualize the degree of conservation of each residue from the amino acid alignment data.
If possible, I'd like to extract the parts of the game that scored ...
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1answer
300 views
How to extract unmatched reads using bwa and samtools?
I have a single read (NOT paired) that I need to pass through the workflow described in Beauclair et al. paper (free version here https://rnajournal.cshlp.org/content/24/10/1285.long) for identifying ...
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299 views
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26 views
which is the right prosite pattern?
I would like to ask if the right Prosite pattern for this multiple alignment:
Is it this:
A-T-[AT]-G-x-C-[AGC]-C-x(1,4)-A
or this:
...
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1answer
148 views
How to calculate mutation rate and mutation sites in a genome using FASTA file?
I have 6 viral genome sequences of the same virus and 1 reference sequence in FASTA format.
How I can identify mutations and mutation sites in those genomes using FASTA sequences but how I can do ...
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75 views
How to change sequence format in my alignment file?
I have fasta file with alingned several sequences (from MUSCLE), when I open it (e.g. notepad++) they look like:
And I want dot format of identities like and then save it in txt file.
I failed ...
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1answer
90 views
Why use Needleman-Wunsch if there is no way to evaluate the statistical significance
I thought that Needleman-Wunsch is the best approach to align sequences. However, I read that it is impossible to evaluate the statistical significance of the alignment if you do global alignment. So ...
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1answer
77 views
merge the matrix from computematrix of deeptools
I have several matrix from computematrix of deeptools. I need to merge two of them using "computeMatrixOperations cbind -m input1.mat.gz input2.mat.gz -o output.mat.gz"
but I am running to error "/...
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98 views
How can I interpret multiple alignment results?
I would like to ask how I can interpret objectively multiple alignment results. I have used JALVIEW and TCOFFEE.
I would like to ask if consistency score plays any role in the interpretation?
I ...
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1answer
114 views
What's the meaning of having different alignments with the same qname in a bam with no duplicates?
I have removed duplicates from my bam file (not secondary, nor suplementary alignments), and then I have looked for some repeated qnames (to assess the failure of a ...
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1answer
103 views
Chromosomal order of supplementary alignment in BAM file "SA" tag
I have several long-read sequences, which when aligned to a human reference genome, aligns to multiple chromosomal fragment as a result of chromothripsis shattering the linear DNA and randomly piecing ...
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2answers
213 views
Extract end position from CIGAR
I was wondering how could I obtain the end position of an alignment using its start position and its CIGAR String.
If an alignment present some soft clipping, for example:
...
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42 views
Unmapping Alternative reads in BAM file
I want to do some HLA typing, most of the tools require the bam file is aligned to primary genome without alternative read handling. From the International Genome Sample Resource project, I can get ...
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1answer
65 views
Translating a genome sequence into possible frames
I have a sequence below from MYC gene about which I need to translate the sequence in all possible frames AND identify (for each frame) which codons are actually used in MYC
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1answer
48 views
Given a multiple sequence alignment, how do I output all of the non-consensus characters by location?
I have an MSA from MAFFT (as a FASTA file; there are several sequences in the MSA), and I would like to quantify the "variants" between these sequences.
What is the standard tool to do ...
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1answer
44 views
choosing a good representative genome subset
I'm trying to build a genomic database for DNA alignments.
I started with NCBI accessions, but the data is very multiplicative, so I want to use subset of [max] N different strains for each specie.
my ...
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1answer
40 views
Running tophat dockerfile in background
I need to use tophat2 to align some sequences, and I wish to use the docker container. These are large sequences so it will take a long time, plus I'm working on a university server so the chances of ...
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3answers
100 views
Make a fasta out of mapped reads without taking into account the reference
I've aligned reads onto an haploid reference genome. I'd like to have a consensus sequence of all my aligned reads that doesn't take into account the reference genome I used (i.e. I just want the ...
2
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1answer
181 views
How can I not show insertions in the Integrative Genome Viewer (IGV)?
I am using IGV 2.5.2 and would not like to see the insertions in my aligned reads. How can I do that? I have managed to remove mismatched bases because there is an option to do so when I right-click ...
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1answer
91 views
Use of Electronic Phenotype in EHR
May I know what's the use of Electronic Phenotyping using EHR data?
I did refer this link but have few questions
I understand that ...
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1answer
72 views
Is there a modern alignment tool tailored for transmembrane regions?
I am looking for a project or tool that allows programmatic pairwise alignments of proteins but that takes care with transmembrane regions of proteins. TM regions are traditionally too information ...
