Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • miaom
    Member
    • Oct 2012
    • 12

    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
  • Torst
    Senior Member
    • Apr 2008
    • 275

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

    Comment

    • mihuzx
      Member
      • Apr 2013
      • 20

      #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

      • rnaeye
        Member
        • May 2011
        • 80

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

        Comment

        Latest Articles

        Collapse

        • 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...
          Today, 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
        • SEQadmin2
          Nine Things a Sample Prep Scientist Thinks About Before Sequencing
          by SEQadmin2


          I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

          Here are nine questions we think about, in roughly the order they matter, before...
          06-18-2026, 07:11 AM

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by SEQadmin2, Today, 10:08 AM
        0 responses
        5 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, Yesterday, 11:05 AM
        0 responses
        7 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 07-02-2026, 11:08 AM
        0 responses
        29 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-30-2026, 05:37 AM
        0 responses
        28 views
        0 reactions
        Last Post SEQadmin2  
        Working...