Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • reason for low mapping rate??

    we did RNASeq using HiSeq 2000 100PE. When the data were back, I mapping them to the reference sequence, but got very low mapping rate (30-40%). I asked around, some people suggest to checked for adaptor contamination.
    I run Trimmomatic and FastQC again. There is almost no improvement. The report and figures are as follows. Could anybody help with the problem?

    #---------------
    PASS Basic Statistics A_1.fq
    PASS Per base sequence quality A_1.fq
    PASS Per sequence quality scores A_1.fq
    PASS Per base sequence content A_1.fq
    PASS Per base GC content A_1.fq
    WARN Per sequence GC content A_1.fq
    PASS Per base N content A_1.fq
    WARN Sequence Length Distribution A_1.fq
    FAIL Sequence Duplication Levels A_1.fq
    WARN Overrepresented sequences A_1.fq
    FAIL Kmer Content A_1.fq
    #---------------
    Figures:
    per_base_quality.png,
    kmer_profiles.png,
    per_sequence_quality.png,
    duplication_levels.png
    .
    Attached Files

  • #2
    DNA or RNA?
    What organism?
    Fresh, frozen, FFPE ?

    Comment


    • #3
      maybe something wrong with the library construction.
      can map the reads to ncbi db or rRNA db to check if the library is pure and no rRNA.
      I am sorry my English is poor.Hope i made it clear.

      Comment


      • #4
        Did you check if unmapped reads mapping to something else such as external contamination etc?

        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