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  • Myrna questions

    Hi all,
    Two small questions about myrna:

    1) I didn't understand how the per gene p-values and q-values are calculated. The p-valuse represent the significance of a gene having a certain gene expression level by the amount of reads mapped to it? By the score of the alignment?

    2) I'm trying to run myrna with bowtie-args "--sam" to get a SAM file of the alignment, and get this error (it works just fine with the same parameters without the flag "bowtie-args").
    Any ides why this happens?

    * sh: line 1: 18915 Segmentation fault /home/sbsuser/bin/bowtie-0.12.7/bowtie --partition 1000000 --mm -t --hadoopout --st
    artverbose --sam --12 .tmp.Align.pl.18662.LOG-sub1-lane8 /data/BKU/Runs/2.7-Dec-10/Lif****z/myrna_dir/jar/index/tomato 2> .tmp.
    Align.pl.18662.LOG-sub1-lane8.err
    * cp * /tmp/error/Align.pl.18662/
    * cp: cannot stat `*': No such file or directory
    * cp .* /tmp/error/Align.pl.18662/
    * cp: omitting directory `.'
    * cp: omitting directory `..'
    * /home/sbsuser/bin/bowtie-0.12.7/bowtie exited with non-zero exitlevel 35584; see /tmp/error/Align.pl.18662 on Pipeline

    Thanks,
    Rachelly.

  • #2
    Dear All,

    Myrna software: http://bowtie-bio.sourceforge.net/myrna/index.shtml
    I am trying to use Myrna software to analyze RNA-seq data from two cell-lines. We repeated experiments twice for each of the cell line hence I merged the data from repeated experiments. However, I am getting an error message while using following command:
    myrna_local --input=Cell1_ Cell2.manifest --preprocess --reference=MYRNA_REFS/human_ensembl_58 --output=output_full --cpus=8 --intermediate=~/intermediate --keep-all --quality=phred64 --family=poisson --tempdir=~/temp

    I am getting following error message:
    Normalization: lup
    Output URL: /gs1/users/rakesh/CIT/Analysis/Myrna/intermediate/count
    Processing label Normal-Subject1-Lane1/2
    stat /gs1/users/rakesh/CIT/Analysis/Myrna/intermediate/count/Normal-Subject1-Lane1/2.txt >& /dev/null
    stat /gs1/users/rakesh/CIT/Analysis/Myrna/intermediate/count/Normal-Subject1-Lane1/2.norm >& /dev/null
    stat /gs1/users/rakesh/CIT/Analysis/Myrna/intermediate/count/Normal-Subject1-Lane1/2.norms >& /dev/null
    mv: cannot move `.tmp.Normal.pl.29153' to `Normal-Subject1-Lane1/2.txt': No such file or directory
    stat /gs1/users/rakesh/CIT/Analysis/Myrna/intermediate/count/2.txt >& /dev/null
    cp Normal-Subject1-Lane1/2.txt /gs1/users/rakesh/CIT/Analysis/Myrna/intermediate/count/ >&2
    cp: cannot stat `Normal-Subject1-Lane1/2.txt': No such file or directory
    Can't load Normal-Subject1-Lane1/2.txt to /gs1/users/rakesh/CIT/Analysis/Myrna/intermediate/count/ (256)

    The Cell1_ Cell2.manifest file details are:
    /data/rakesh/CIT/Analysis/Myrna/Cell1_F_combined.txt 0 /data/rakesh/CIT/Analysis/Myrna/Cell1_R_combined.txt 0 Normal-Subject1-Lane1
    /data/rakesh/CIT/Analysis/Myrna/Cell2_F_combined.txt 0 /data/rakesh/CIT/Analysis/Myrna/Cell2_R_combined.txt 0 Tumor-Subject1-Lane1

    Can you please help me to solve the issue?

    I really appreciate your help,

    Thanks,

    Rakesh

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