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  • 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

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