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  • cz0013
    Junior Member
    • Jul 2014
    • 8

    Bowtie2-build error writing to reference index file

    I have been trying to make an index file for bowtie2.

    What I type is:
    bowtie2-build sequence.fasta sequence

    Then I get this error:
    Settings:
    Output files: "sequence.*.bt2"
    Line rate: 6 (line is 64 bytes)
    Lines per side: 1 (side is 64 bytes)
    Offset rate: 4 (one in 16)
    FTable chars: 10
    Strings: unpacked
    Max bucket size: default
    Max bucket size, sqrt multiplier: default
    Max bucket size, len divisor: 4
    Difference-cover sample period: 1024
    Endianness: little
    Actual local endianness: little
    Sanity checking: disabled
    Assertions: disabled
    Random seed: 0
    Sizeofs: void*:8, int:4, long:8, size_t:8
    Input files DNA, FASTA:
    sequence.fasta
    Building a SMALL index
    Reading reference sizes
    Error writing to the reference index file (.4.ebwt)
    Time reading reference sizes: 00:00:01
    Total time for call to driver() for forward index: 00:00:01
    Error: Encountered internal Bowtie 2 exception (#1)
    Command: bowtie2-build --wrapper basic-0 sequence.fasta sequence
    Deleting "sequence.3.bt2" file written during aborted indexing attempt.
    Deleting "sequence.4.bt2" file written during aborted indexing attempt.

    If anyone knows what is causing this error help would be greatly appreciated.
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

    #2
    That is a bit odd since you seem to have write permissions in the directory where you are running this command (is that correct)? What kind (length, multifasta)) of reference is in sequence.fasta?

    Are you using the latest bowtie2 package?

    Comment

    • cz0013
      Junior Member
      • Jul 2014
      • 8

      #3
      I do have write permissions, and I should be using the latest bowtie2 package. sequence.fasta is the the SL2.5 release of the tomato genome.

      Comment

      • GenoMax
        Senior Member
        • Feb 2008
        • 7142

        #4
        Can you try using a different "base name" for the index files (e.g. tomato instead of sequence)? One additional thing to try would be to force bowtie to create a "large" index by adding --large-index to command line.

        Comment

        • cz0013
          Junior Member
          • Jul 2014
          • 8

          #5
          I am now running like this:
          bowtie2 --large-index SL2.50_genome.fasta SL2.50_genome

          But now I get this error message:
          bowtie2 --large-index SL2.50_genome.fasta SL2.50_genome
          (ERR): Cannot find the large index 0.1.bt2l
          Exiting now ...

          Comment

          • GenoMax
            Senior Member
            • Feb 2008
            • 7142

            #6
            You need to do "bowtie2-build" first before you can use the index for alignments with bowtie2. Did the indexing part work?

            Are you now ready to do the alignment (looks like it)?

            Comment

            • cz0013
              Junior Member
              • Jul 2014
              • 8

              #7
              That is my bad. I should have been running bowtie2-build. I ran bowtie2 without realizing that it was the wrong command.I ran it again this time actually using bowtie2-build, and I ran into the same problem that I was having before. So I haven't gotten the indexing part to work.
              Settings:
              Output files: "SL2.50_genome.*.bt2l"
              Line rate: 7 (line is 128 bytes)
              Lines per side: 1 (side is 128 bytes)
              Offset rate: 4 (one in 16)
              FTable chars: 10
              Strings: unpacked
              Max bucket size: default
              Max bucket size, sqrt multiplier: default
              Max bucket size, len divisor: 4
              Difference-cover sample period: 1024
              Endianness: little
              Actual local endianness: little
              Sanity checking: disabled
              Assertions: disabled
              Random seed: 0
              Sizeofs: void*:8, int:4, long:8, size_t:8
              Input files DNA, FASTA:
              SL2.50_genome.fasta
              Building a LARGE index
              Reading reference sizes
              Error writing to the reference index file (.4.ebwt)
              Time reading reference sizes: 00:00:01
              Total time for call to driver() for forward index: 00:00:01
              Error: Encountered internal Bowtie 2 exception (#1)
              Command: bowtie2-build --wrapper basic-0 SL2.50_genome.fasta SL2.50_genome
              Deleting "SL2.50_genome.3.bt2l" file written during aborted indexing attempt.
              Deleting "SL2.50_genome.4.bt2l" file written during aborted indexing attempt.

              Comment

              • westerman
                Rick Westerman
                • Jun 2008
                • 1104

                #8
                Is it possible that you are running out of disk space? A quick and dirty way of checking is to start up the indexing process and in another window do something like:

                for i in `seq 1 20`; do df -h; sleep 60

                That way you will get 20 snapshots of disk space usage at 1 minute intervals. You should see the disk usage go up one one of your disks but, we hope, not to the limit.

                Comment

                • cz0013
                  Junior Member
                  • Jul 2014
                  • 8

                  #9
                  I think that was the problem. The server that I was running it on is out of space. I switched servers and now it is working. Thanks for the help guys, much appreciated.

                  Comment

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