Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Help with plotting ngs (small RNA seq) data

    Hello all,

    I am new to ngs (an unfortunately to bioinfo as well) , I Recently sequenced small RNAs (21-29 nt) using Illunina platform. I aligned the reads on to genome using bowtie, and extracted alignments for regions of interest with samtools, I am normalizing data among the samples by counting rRNA reads.

    I want to now to count the reads (total number) originating from certain region of genome and plot the read density (for both + and -strands together). Please suggest me any Rpackage or any other suitable method for counting reads and plotting them.
    Last edited by Ami; 06-01-2011, 09:00 AM.

  • #2
    I would:
    1. use bedtools to convert bam to a bedgraph file
    2. convert the bedgraph to bigwig (ucsc utility)
    3. place bigwig on a web server
    4. point UCSC genome browser at bigwig file

    Comment


    • #3
      You could use:

      Galaxy (web based tools, http://main.g2.bx.psu.edu/)

      Integrated Genome Viewer (get .sam format from bowtie, convert to .bam using samtools, use coverage tool in IGV and also plot it there),

      SeqMonk (another integrated software that can do this),

      Artemis (may be somewhat confusing to learn initially, but have some flexible plotting tools (using 'User graphs' option),

      R bioconductor for sequence analysis (http://www.bioconductor.org/help/wor...ut-sequencing/)

      Comment


      • #4
        I use bowtie -> Sam -> Bam -> IGV

        Comment


        • #5
          Thank you everybody for the help, I am trying the suggested options to find out which is easy for me.

          Comment

          Latest Articles

          Collapse

          • 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
          • seqadmin
            Strategies for Sequencing Challenging Samples
            by seqadmin


            Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
            03-22-2024, 06:39 AM

          ad_right_rmr

          Collapse

          News

          Collapse

          Topics Statistics Last Post
          Started by seqadmin, 04-11-2024, 12:08 PM
          0 responses
          18 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 10:19 PM
          0 responses
          22 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 09:21 AM
          0 responses
          16 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-04-2024, 09:00 AM
          0 responses
          47 views
          0 likes
          Last Post seqadmin  
          Working...
          X