Are you using latest tophat? Are you able to run TopHat with just the -G option without providing reads (so the transcriptome index can be pre-built?)
From TopHat manual
For example, in order to just prepare the transcriptome index files for a specific annotation, an initial, single TopHat run could be invoked like this:
tophat -G known_genes.gtf \
--transcriptome-index=transcriptome_data/known \
hg19
In this example TopHat will create the transcriptome_data directory in the current directory (if it doesn't exist already) containing files known.gff, known.fa, known.fa.tlst, known.fa.ver and the known.* Bowtie index files. Then for subsequent TopHat runs with the same genome and known transcripts but different reads, TopHat will no longer spend time building the transcriptome index because it can use the previously built transcriptome index files, so the -G option is no longer needed (however using it again will not force TopHat to rebuild the transcriptome index files if they are already present and with the matching version)
tophat -G known_genes.gtf \
--transcriptome-index=transcriptome_data/known \
hg19
In this example TopHat will create the transcriptome_data directory in the current directory (if it doesn't exist already) containing files known.gff, known.fa, known.fa.tlst, known.fa.ver and the known.* Bowtie index files. Then for subsequent TopHat runs with the same genome and known transcripts but different reads, TopHat will no longer spend time building the transcriptome index because it can use the previously built transcriptome index files, so the -G option is no longer needed (however using it again will not force TopHat to rebuild the transcriptome index files if they are already present and with the matching version)
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