How to run a Alternative Splice Junction analysis ?
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5.8 years ago

Hello everyone, I am totally new to bioinformatics and programming, but i'm sure it's the future of science and I am determined to learn .

So I will try to explain my problem, and maybe someone can help me. I have RNA-seq data, already processed and analyzed. So I have bam/bai files in a directory, and excel files with readable results of diff. expressed genes between my two conditions.

I found a program on Git Hub, name spladder https://github.com/ratschlab/spladder which should do more or less what I want, i.e find if there is differents splicing variants between my two conditions. Maybe there is better tools, if someone knows one...

The problem is that I can't even run the program, it must be something stupid but i'm totally new, and this is a nightmare for me. I'm running on Mac OS, latest version (mojave). I've installed anaconda3 on my mac, and I have a directory name anaconda3 in my harddrive, along with user, system etc.

I used git clone on my terminal to clone the repository of spladder on github. It appears in a depository name projects on my "user" page , with downloads, documents etc.

When i try to run python spladder.py in terminal, I have : :~ user$ python3 spladder.py python3: can't open file 'spladder.py': [Errno 2] No such file or directory

my PATH is user$ echo $PATH /anaconda3/bin:/miniconda3/bin:/usr/local/bin:/usr/bin:/bin:/usr/sbin:/sbin

I tried to copy/past the "projects" directory in anaconda3 file, but it changed nothing. I think there is a very simple thing I miss here...

I hope someone can help, with a lot of gratitude, Thanks

python mac Alternative splicing RNA-Seq • 1.5k views
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Does python /path/to/where/you/git/cloned/spladder/python/spladder.py work?

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4.4 years ago

Hi Dear, I just successfully Run same as your problem.. I am not sure that may I explain you well or not... but I try...

You need to use samtools package for indexing your bam file second, you also need to create an annotation file form gencode.v25ERCC.annotation.fixedchrMT.gtf to gencode.v25ERCC.annotation.fixedchrMT.gtf.pickle

However, you don't use these index and gtf.pickle file but these must already exist in your folder. then you run.

My successful code is

spladder build -b /home/jabbar/projects/BamStar/ANTINS_113_alts/Aligned.sortedByCoord.out.bam -o /home/jabbar/spladder -a /home/jabbar/projects/RNAseq/annotation/annotation/gencode.v25ERCC.annotation.fixedchrMT.gtf

If you have any other issue than this or in this you may ask .

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