As I have stated above, I have received an error in running tophat, with the output below. The command I supplied was tophat humanGRCh38 gerald_C2DBEACXX_3.fq,gerald_C1TD1ACXX_8_ACAGTG.fq , as I was running tophat from the directory in which the bt2l files were installed. I have supplied the prefix as well, but it did not work. I have gotten the same error when I supply the entire present working directory and the prefix, I got the same problem. Anyone else run into this, and if yes how did you fix it? I know that the version of bowtie that I am using is correct because the files are in the correct format.
$ ls
gerald_C1TD1ACXX_8_ACAGTG.bam Homo_sapiens.GRCh38.96.gtf humanGRCh38.3.bt2l
gerald_C1TD1ACXX_8_ACAGTG.fq Homo_sapiens.GRCh38.dna.toplevel.fa humanGRCh38.4.bt2l
gerald_C2DBEACXX_3.bam humanGRCh38.1.bt2l tophat_out gerald_C2DBEACXX_3.fq humanGRCh38.2.bt2l
$ tophat humanGRCh38 gerald_C2DBEACXX_3.fq,gerald_C1TD1ACXX_8_ACAGTG.fq
[2019-07-02 08:34:27] Beginning TopHat run (v2.1.1)
-----------------------------------------------
[2019-07-02 08:34:27] Checking for Bowtie
Bowtie version: 2.3.4.1
[2019-07-02 08:34:27] Checking for Bowtie index files (genome)..
Error: Could not find Bowtie 2 index files (humanGRCh38.*.bt2l)
$
Did you make this index yourself? Has it been tested before this?
What do you get if you do:
You should see the names of chromosomes being listed like this (truncated, may not look exactly like this, we are testing to make sure you don't get an error).
No, I did not build this index myself. I used a pre-built index from ftp://ftp.ncbi.nlm.nih.gov/genomes/archive/old_genbank/Eukaryotes/vertebrates_mammals/Homo_sapiens/GRCh38/seqs_for_alignment_pipelines/GCA_000001405.15_GRCh38_no_alt_analysis_set.fna.bowtie_index.tar.gz
When I do bowtie2-inspect -n humanGRCh38 I get an error
I see the following files in the tar archive you linked above. So it looks like you did not get the full set of files or uncompressing went wrong somewhere.
Unless absolutely essential you may want consider using a current RNAseq splice aware aligner like
STAR
,bbmap
instead ofTopHat
.