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  1. IGB
  2. IGBF-3165

Make junction files for experimental and positive control alignments data

    Details

    • Type: Task
    • Status: Closed (View Workflow)
    • Priority: Major
    • Resolution: Done
    • Affects Version/s: None
    • Fix Version/s: None
    • Labels:
      None

      Description

      Run the "find junctions" program to generate spreadsheets showing support for splicing events.

      This program generates bed format files that tabulate evidence supporting splicing. The "score" field of the output file indicates the number sequence alignments that exhibit evidence of splicing in a particular location, called a "junction".

      This format for representing splicing events comes originally from the "tophat" program, which generates "bed" files containing junctions in essentially the same way.

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            Hide
            ann.loraine Ann Loraine added a comment - - edited

            Added new files to hotpollen/splicing_analysis repository:

            • find_junctions.sh
            • sbatch-doIt.sh
            • find-junctions "jar" file

            Checked that the system has java:

            [aloraine@str-i1 bams]$ java -version
            openjdk version "1.8.0_342"
            OpenJDK Runtime Environment (build 1.8.0_342-b07)
            OpenJDK 64-Bit Server VM (build 25.342-b07, mixed mode)
            

            Created new directory "bams" with symbolic links to all the BAM files and their indexes made in IGBF-3143.
            Made symbolic links to above scripts and jar file within this location.
            Launched jobs with:

            sbatch-doIt.sh .bam find_junctions.sh >jobs.out 2>jobs.err
            

            Got error:

            Error: Invalid or corrupt jarfile find-junctions-1.0.0-jar-with-dependencies.jar

            Fixed it by copying a fresher version of the jar file to the "src" directory in my home folder.

            Show
            ann.loraine Ann Loraine added a comment - - edited Added new files to hotpollen/splicing_analysis repository: find_junctions.sh sbatch-doIt.sh find-junctions "jar" file Checked that the system has java: [aloraine@str-i1 bams]$ java -version openjdk version "1.8.0_342" OpenJDK Runtime Environment (build 1.8.0_342-b07) OpenJDK 64-Bit Server VM (build 25.342-b07, mixed mode) Created new directory "bams" with symbolic links to all the BAM files and their indexes made in IGBF-3143 . Made symbolic links to above scripts and jar file within this location. Launched jobs with: sbatch-doIt.sh .bam find_junctions.sh >jobs.out 2>jobs.err Got error: Error: Invalid or corrupt jarfile find-junctions-1.0.0-jar-with-dependencies.jar Fixed it by copying a fresher version of the jar file to the "src" directory in my home folder.
            Hide
            ann.loraine Ann Loraine added a comment -

            Forgot to include the 2bit file in the directory. Adding it and restarting the jobs.

            Show
            ann.loraine Ann Loraine added a comment - Forgot to include the 2bit file in the directory. Adding it and restarting the jobs.
            Hide
            ann.loraine Ann Loraine added a comment -

            Jobs look done, with 66 ".tbi" index files, one per each of the 66 input .bam files.

            Show
            ann.loraine Ann Loraine added a comment - Jobs look done, with 66 ".tbi" index files, one per each of the 66 input .bam files.
            Hide
            ann.loraine Ann Loraine added a comment -

            Moved files to:

            /nobackup/tomato_genome/alt_splicing/for_igbquickload/junction_files

            Show
            ann.loraine Ann Loraine added a comment - Moved files to: /nobackup/tomato_genome/alt_splicing/for_igbquickload/junction_files
            Hide
            ann.loraine Ann Loraine added a comment - - edited

            To review:

            • check that files have reasonable sizes (no "zero" size files, for example)
            • check that every "FJ.bed.gz" file has a corresponding "FJ.bed.gz.tbi" index file
            • check that every "SRR" bam file in our control and experimental sample directories has a corresponding "FJ.bed.gz" file
            Show
            ann.loraine Ann Loraine added a comment - - edited To review: check that files have reasonable sizes (no "zero" size files, for example) check that every "FJ.bed.gz" file has a corresponding "FJ.bed.gz.tbi" index file check that every "SRR" bam file in our control and experimental sample directories has a corresponding "FJ.bed.gz" file
            Hide
            Mdavis4290 Molly Davis added a comment -

            Review:

            Directory- /nobackup/tomato_genome/alt_splicing/for_igbquickload/junction_files

            • No files were zero size. Used command LL/ll.
            • Every "FJ.bed.gz" file has a corresponding "FJ.bed.gz.tbi" index file
            • Every "SRR" bam file in our control and experimental sample directories has a corresponding "FJ.bed.gz" file. SRP328042-molly SRR files are in-between SRP252265 files in the directory might be hard to differentiate which is control and which is experimental.

            [~aloraine]

            Show
            Mdavis4290 Molly Davis added a comment - Review: Directory- /nobackup/tomato_genome/alt_splicing/for_igbquickload/junction_files No files were zero size. Used command LL/ll. Every "FJ.bed.gz" file has a corresponding "FJ.bed.gz.tbi" index file Every "SRR" bam file in our control and experimental sample directories has a corresponding "FJ.bed.gz" file. SRP328042-molly SRR files are in-between SRP252265 files in the directory might be hard to differentiate which is control and which is experimental. [~aloraine]
            Hide
            ann.loraine Ann Loraine added a comment -

            Thanks [~molly]. I decided to store all the files in the same folder and will use an index / table of contents type strategy to distinguish them. Moving to Done.

            Show
            ann.loraine Ann Loraine added a comment - Thanks [~molly] . I decided to store all the files in the same folder and will use an index / table of contents type strategy to distinguish them. Moving to Done.

              People

              • Assignee:
                ann.loraine Ann Loraine
                Reporter:
                ann.loraine Ann Loraine
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                • Created:
                  Updated:
                  Resolved: