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  • Ami
    Junior Member
    • Aug 2010
    • 6

    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.
  • adamdeluca
    Member
    • Jul 2010
    • 95

    #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

    • Kennels
      Senior Member
      • Feb 2011
      • 149

      #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

      • NicoBxl
        not just another member
        • Aug 2010
        • 264

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

        Comment

        • Ami
          Junior Member
          • Aug 2010
          • 6

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

          Comment

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