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Case Study 1. exRNA-seq

Basic analyses for exRNA-seq

a) Background Introduction

  • PPT: 0. Introduction of exRNA-seq.pdf (view on-line only, not downloadable)
  • Teaching Video: Week V - Part II. 0. Intr to exRNA-seq.mov

b) Understand your data

Type of RNA-seq

  • Default: (whole cell poly-A) RNA-seq (>200nt)
  • Other types:
    • small RNA-seq (<50nt)
    • total RNA-seq (ribosome removed) (>200nt)
    • nonpolyA RNA-seq (ribosome removed) (>200nt)
  • Different cell localizations:
    • nuc. (total)
    • chromosome (total)
    • cyto. (poly-A)
  • Single cell RNA-seq
  • exRNA-seq
    • cell free/MV/exosome/RNP
    • small/long

So what RNA-seq we are mapping and analyzing?

  • Sequencing machine ?
  • Single-strand V.S. Paired-end ?
  • Strand specific ?
  • Size selection ?
  • Poly-A enriched or total (ribosome removed) ?
  • Cellular localization ?

c) Organize your data

Data format:

  • fasta
  • fastaq
  • gff/gtf
  • bam
  • bed
  • bigwig

Shared dirs/data:

cd  #go to my home
ln -s /BioII/lulab_b/shared/genomes  .   #shared reference genome sequeunces and annotations
ln -s /BioII/lulab_b/shared/shared_scripts  .  #shared scripts in Lu Lab
ln -s /BioII/lulab_b/shared/projects/exRNA  shared_exRNA_projects   #shared projects' files
  • ~/genomes/human_hg38
    • sequences/ #sequences of reference genome (fasta format)
    • index/ # indexed genome sequences
    • gtf/ #annotation of reference genome

Make your own project dir:

cd # go to my home
vim .bashrc # see my example in /home/john/.bashrc (and /home/john/shortcuts)
mkdir github

mkdir -p ~/projects/exRNA
cd ~/projects/exRNA

alias mkpr="mkdir -p {data/mapped,scripts,analysis}"  # you can put this in your ~/.bashrc
mkpr

d) Get the software ready

Install bioinformatics software in Linux (centos)

Video

@Youtube

@Bilibili

Other pipelines for RNA-seq analyses

{% hint style="success" %} We also recommend some other Tutorials/Pipelines you can learn from:

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