Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • smallSeq
    Junior Member
    • Nov 2014
    • 6

    Galaxy smallRNA Analysis help

    Hi,
    I've been playing with Galaxy lately to do my data analysis for smallRNA (Illumina).
    I think I'm getting what I want but I'm stuck at a certain point.
    Here are the tools/workflow I have used:
    1. FastQC (to check quality)
    2. Clip (to trim off 3' adapter sequence)
    3. Sickle (to discard any bad quality sequences)
    4. MiRDeep2 mapper to collapse the reads from Sickle (seems to just make the sickle file compatible with MiRDeep2 Quantifier)
    5. MiRDeep 2 Quantifier to map the collapsed reads to the reference fasta file of mature miRNA.
    *Result is a list of miRNA in a tabular format
    -Next I want to convert this file to a file compatible with edgeR for differential expression analysis. Any thoughts on how to do this? Any known tools?
    -Also, there is one other tool: MiRDeep2 (identification of novel and known miRNAs). I can't for the life of me figure out if this tool is more useful or if I should be using this one instead of Quantifier however the output when I use this tool is not giving me the miRNA list I would expect. Any help would be appreciated. Thx!!
  • Schelarina
    Member
    • Apr 2014
    • 18

    #2
    hi,
    does mirdeep provide you with a list of counts for each miRNA? if not, i guess it gives at least the genomic coordinates of your miRNAs, so you can make it as bedfile or gff file and use it to extract read counts with betools or other tools on your bam file. The bed file you need it anyway for edgeR
    Then you just extract the coloumn with the counts in a new txt file and give to edgeR lie this
    counts <- read.delim("counts.txt")

    Comment

    • smallSeq
      Junior Member
      • Nov 2014
      • 6

      #3
      Thanks for your reply. I'm just getting back to trying to figure this out.
      I get a list of counts (see attached image) for each miRNA when mapping to the mature fasta file downloaded from miRbase. These aren't necessarily assigned to regions in the genome however, just miRNA names. This is a tabular file. It seems like edgeR wants me to combine all my lists(samples and replicates) together into a matrix type tabular file. Do you know of a tool to do this easily? Will edgeR accept these counts if not assigned to a position in the genome and just a name? Any help is appreciated. Thx!
      Attached Files

      Comment

      Latest Articles

      Collapse

      • SEQadmin2
        Advanced Sequencing Platforms Tackle Neuroscience’s Toughest Genomics Problems
        by SEQadmin2



        Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
        ...
        07-09-2026, 11:10 AM
      • SEQadmin2
        Cancer Drug Resistance: The Lingering Barrier to Rising Survival
        by SEQadmin2



        Cancer survival rates have significantly increased in the last few decades in the United States, reaching a combined 70% 5-year survival rate by 2021. Behind this number, there are years of research to find new therapies, drug targets, and early detection methods. But there is one core challenge that keeps slowing down these advances, and it’s about drug resistance.

        There is no single reason why many patients don’t respond to treatment as expected. Cancer is...
        07-08-2026, 05:17 AM
      • GATTACAT
        Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
        by GATTACAT
        Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
        07-01-2026, 11:43 AM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by SEQadmin2, 07-09-2026, 10:04 AM
      0 responses
      23 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 07-08-2026, 10:08 AM
      0 responses
      15 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 07-07-2026, 11:05 AM
      0 responses
      33 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 07-02-2026, 11:08 AM
      0 responses
      31 views
      0 reactions
      Last Post SEQadmin2  
      Working...