UAB Galaxy RNA Seq Step by Step Tutorial
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Galaxy RNA-Seq Step-by-Step Tutorial/Protocol
Upload data
For this tutorial, we will have 2 samples, sequences with paired-end reads. That gives us 4 FASTQ files to upload.
set filetype and genome
To start, you must move the data (FASTQ) from the sequencing center into the Galaxy instance, be sure to specify the filetype (fastqsanger for UAB and HudsonAlpha) and the organism that was sequenced ("Genome database"), in this tutorial, "mm9" for mouse.
check quality of data
At this step, we check the quality of sequencing. This says nothing about the quality of the sample, or whether it was the right sample. We'll check that later.
. This
Align using TopHat
QC alignment
* %mapped * visualize in IGV or IGB
Construct transcripts : Cufflinks
* Denovo vs existing annotation * UAB Modified Reference annotations