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  • Question: Cufflinks displaying "WARNING: Multi-Hit not found"

    I'm running Cufflinks (2.2.1) on a set of RNA-seq alignments that I did with STAR. I'm using these options:

    Code:
    /home/user1/Data/Softwares/cufflinks-2.2.1/cufflinks -p 6 -u -o ./Cufflinks/ERR036348_pass-2 ./Alignments/ERR036348_pass-2/ERR036348_pass-2_Aligned.sortedByCoord.out.bam
    In every instance I get the message: "WARNING: Multi-Hit not found". I pasted the beggining of the output of a run I did.

    Code:
    [00:12:17] Inspecting reads and determining fragment length distribution.
    > Processed 51509 loci.                        [*************************] 100%
    > Map Properties:
    >       Normalized Map Mass: 28150478.05
    >       Raw Map Mass: 28150478.05
    >       Number of Multi-Reads: 2479202 (with 6180369 total hits)
    >       Fragment Length Distribution: Empirical (learned)
    >                     Estimated Mean: 353.78
    >                  Estimated Std Dev: 35.57
    [01:36:09] Assembling transcripts and initializing abundances for multi-read correction.
    > Processing Locus 1:86701401-87057627         [                         ]   2%
    WARNING: Multi-Hit not found (86357258,86357334).
    
    WARNING: Multi-Hit not found (86357265,86357341).
    
    WARNING: Multi-Hit not found (86357276,86357352).
    In this particular library, I got the warning 1997 times.

    What is happening? Should I add a option to Cufflinks?

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