Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • Rsubread for miRNA

    For micro RNA (miRNA) data, the following aligners are recommended specifically for these short sequences:
    MicroRazerS (www.seqan.de/projects/microrazers/)
    mrFAST (mrfast.sourceforge.net/)
    mrsFAST (mrsfast.sourceforge.net/Home)
    PatMaN (bioinf.eva.mpg.de/patman/)
    Does anyone know how the Rsubread align function compares to these? Has anyone performed any comparisons? I use Rsubread for RNAseq and it would be convenient to use it also for miRNAseq, but I am a little concerned and wonder whether I need to invest time in conducting some comparisons.

    I have noticed one potential problem with Rsubread featureCounts function when applied to miRNAseq: When I use the annotation file from mirBase (hsa.gff3) instead of the built-in annotation or the ensembl GTF file, then the Gene IDs in the counts (rownames) and annotation output from Rsubread-featureCounts are all NA (see code below).

    counts_TH14_uniqtrue_annotMirBmature.out <- featureCounts(files=mapped.flist,
    annot.inbuilt="hg38", chrAliases=NULL,
    # use mirBase GTF file and feature = miRNA (mature miRNA)
    annot.ext="/home/inah/Rsubread_miRNA/RefGTF/hsa.gff3",
    isGTFAnnotationFile=TRUE,
    GTF.featureType="miRNA", GTF.attrType="miRNA", useMetaFeatures=FALSE, ...

    Many thanks, Ina

  • #2
    For miRNA I used bowtie1 with --best --strata and bowtie2 with --very-sensitive-local options. I found around a 90% alignment score between my reads and reference genome.

    Based on Ziemann et al 2016. "Evaluation of miRNA alignment techniques".

    This is the code I used for featureCounts:

    featureCounts -t miRNA -g ID -f -s 1 -O -T 16 -a /smallRNA.gtf -o /output_dir n.bam 2>featureCounts.log

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Essential Discoveries and Tools in Epitranscriptomics
      by seqadmin


      The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist on Modified Bases...
      Yesterday, 07:01 AM
    • seqadmin
      Current Approaches to Protein Sequencing
      by seqadmin


      Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
      04-04-2024, 04:25 PM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, 04-11-2024, 12:08 PM
    0 responses
    39 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 10:19 PM
    0 responses
    41 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 09:21 AM
    0 responses
    35 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-04-2024, 09:00 AM
    0 responses
    55 views
    0 likes
    Last Post seqadmin  
    Working...
    X