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  • Can pre-filtering reads affect your analysis results?

    I tried to run TopHat/Cufflinks using Illumina reads without and without quality-filtering. I noticed that the results of differential expression analysis are significantly different. For example, a gene that was highly DE when reads were not pre-filtered, was absolutely unchanged when reads were pre-filtered.

    Why would the removal of BAD quality reads affect the results so dramatically?

    Thanks!

    Note: prefiltering = remove the pairs where one of the mate has all bases with quality < 2.

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