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6.8 years ago
Whoknows
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960
Hello
I have only one human RNA-SEQ sample and I wanted to find novel isoforms and also novel genes from this sample.
I have performed Topaht2 and Cufflinks, which cufflinks generates 4 files for me.
Should I run Cuffcompare to find novel isoforms / genes? or I should use cufflinks output files?
Thanks
You should know that the old 'Tuxedo' pipeline of Tophat and Cufflinks is no longer the "advisable" tool for RNA-seq analysis. The software is deprecated/ in low maintenance and should be replaced by HISAT2, StringTie and ballgown. See this paper: Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown. (If you can't get access to that publication, let me know and I'll -cough- help you.) There are also other alternatives, including alignment with STAR and bbmap, or pseudo-alignment using kallisto or salmon.
thanks, I have read that 2016 paper but I cannot find relevant information about how to find novel isoforms by HISAT. I just want to quantify expression of novel isoforms.