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  • cuffdiff- no assembled transcripts

    Hi

    We are running tophat pipeline on algal sequences and our cuffdiff output does not appear to have assembled transcripts listed, rather CDS that correlate w/ single exons only.

    tophat:
    tophat2 -o ./psr1_rep_s2 -i 50 -I 28000 --min-segment-intron 50 --max-segment-intron 12800 -G Creinhardtii_236_gene_exons.gtf $BOWTIE_INDEX/bowtie2/cre5.3 psr1_p_replete_s2.fastqsanger.gz

    cufflinks:
    cufflinks -o ./cufflinks_out_psr1_rep_s2 -I 50000 -N –g Creinhardtii_236_gene_exons.gtf tophat_out_psr1_rep_s2/accepted_hits.bam

    cuffdiff:
    cuffdiff -o ./cuffdiff_out -L 1690_rep,psr1_rep,1690_dep,psr1_dep /merged_asm/merged.gtf list of accepted_hits.bam


    example cuffdiff which returns tests for DE for each exon, but not entire transcript:
    XLOC_000003 XLOC_000003 PAC:27578295.CDS.1 chromosome_1:20355-20687
    XLOC_000004 XLOC_000004 PAC:27578295.CDS.2 chromosome_1:20759-21065
    XLOC_000005 XLOC_000005 PAC:27578295.CDS.3 chromosome_1:21137-21443
    XLOC_000006 XLOC_000006 PAC:27578295.CDS.4 chromosome_1:21513-21819
    XLOC_000007 XLOC_000007 PAC:27578295.CDS.5 chromosome_1:21986-22132
    XLOC_000008 XLOC_000008 PAC:27578295.CDS.6 chromosome_1:22453-22699
    XLOC_000009 XLOC_000009 PAC:27578295.exon.7 chromosome_1:22862-22948
    XLOC_000010 XLOC_000010 PAC:27578295.exon.8 chromosome_1:23106-23250
    XLOC_000011 XLOC_000011 PAC:27578295.exon.9 chromosome_1:23370-23957


    What am I missing besides the obvious?

    thanks

    Charles

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