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problem in terminal exon assembly using cufflinks

Hello,

I used Cufflinks (v.2.0.2) to assemble a complete transcriptome without providing any GTF references just to check the quality of assembly and it seemed to work so well, but I am worried several terminal exons were not entirely reconstructed (see attached figures in which I compared my cufflinks results with RefSeq annotations using IGV browser) although read coverage was quite good. I tried to change several parameters (disabling low-coverage 3' trimming; decreasing minimum reads per transfrags, min-isoform-fraction, or small-anchor-fraction) but the problem persisted.

Did somebody have the same problem or know how to fix it?

Thanks, Jorge

rna-seq assembly coverage cufflinks

1 answer

Hi everyone,

I realized that it is a problem of Cufflinks version (v.2.0.2). Using newer versions, terminal exons are correctly assembled.

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