RNA-seq analysis using R
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@justinkablan225-18560
Last seen 4.9 years ago

Hello all,
I'm a student and a beginer with R tool for RNA-seq analysis.
I've some Fastq files that I want to (i) convert into BAM file using LIMMA package in R and (ii) make an alignment with genome reference using Toophat tool.

The probleme is that, after reading the LIMMA userguide, I didn't catch what scripts use for those preliminary analysis.

May you help me if possible.

Sincerly.

limma rnaseq convert R • 1.4k views
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Please only tag relevant packages as this emails the package maintainer. I removed the DESeq2 tag.

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@gordon-smyth
Last seen 2 hours ago
WEHI, Melbourne, Australia

The Rsubread package maps reads to a reference genome (which is how you convert FASTQ files into BAM). See

https://www.biorxiv.org/content/early/2018/07/26/377762

or

https://subread.sourceforge.net/RNAseqCaseStudy.html

TopHat is obsolete now; even the TopHat authors themselves recommend that you use newer tools.

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@james-w-macdonald-5106
Last seen 1 hour ago
United States
  1. limma isn't designed to convert FASTQ files into BAM files. After reading (?) the limma User's Guide, I am mystified as to why you would think that it is intended for that purpose. Can you say why you think that?
  2. TopHat isn't a Bioconductor tool, and this is a forum to help people use Bioconductor tools only. You could post your question about TopHat on Biostars.org, or better yet just read the TopHat manual. You will get much farther by doing your own learning rather than simply trying to get others to write a tutorial for you.
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Hi

I done some mistakes in my message but now I begin to understand better more.

Thus, I'm a Bioconductor tool user.

Best regard

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