Hi,

I'm attempting RNA-seq data analysis on Vibrio cholerae samples, at galaxy main https://main.g2.bx.psu.edu/.

I was able to align transcripts using Ion Torrent's TMAP aligner. I uploaded the BAM files and used the cufflinks tool, which gave me FPKM scores for various transcripts. Now I want to extract the relevant gene_id for each mapped transcript using a reference genome. I tried using CuffCompare for this purpose using a reference file from the UCSC archaea database or using a FASTA file from history. In either case, cuffcompare gives me an error. 

Error1: (When using a reference genome from the UCSC archaea database)
 An error occurred running this job:cuffcompare v2.0.2 (3522) No sequence data found for dbkey vibrChol1, so sequence data cannot be used. cp: cannot stat `/galaxy/main_pool/pool4/tmp/job_working_directory/004/846/4846005/cc_output.input1.refmap': No such file or directory cp: cannot stat `

Error 2: (When using a reference genome from history)
An error occurred running this job:cuffcompare v2.0.2 (3522) cuffcompare -o cc_output -r /galaxy/main_pool/pool1/files/005/515/dataset_5515739.dat -s ref.fa ./input1 Error running cuffcompare. You are using Cufflinks v2.0.2, which is the most recent release. No fasta index found for ref

Could you help me figure out what's going on?

Thanks,
-Samit
 
Regards,

Samit Watve
Graduate Teaching Assistant,
School of Biology, Georgia Institute of Technology

PhD Student at the Hammer Lab, 
Office: Cherry Emerson, 218-E, 310 Ferst Drive Atlanta GA
School of Biology, Georgia Institute of Technology
Lab Page: http://www.hammerlab.biology.gatech.edu/