Rui:RNAseq analysis on Hiseq111005: Difference between revisions

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>RuiLiu
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* Lesson 1: Use exactly same VERSION of programs/softwares to do analysis on ALL data in ONE computer
* Lesson 1: Use exactly same VERSION of programs/softwares to do analysis on ALL data in ONE computer
* Lesson 2: Random check for comparison
* Lesson 2: Random check for comparison
*[[Rui:Tophat_11.10.11|Tophat_11.10.11]]
*[[Rui:Cufflinks_11.11.11|Cufflinks_11.11.11]]
*[[Rui:Cuffdiff_11.11.11|Cuffdiff_11.11.11]]


===Tophat_11.10.11===
===Tophat_11.10.11===

Revision as of 01:59, 22 November 2011

RNAseq analysis on Hiseq111005

Comparison of program

' ' Meangenemachine Genome-miner ' '
Tophat v1.3.1 v1.3.1
Bowtie 0.12.7.0 0.12.7.0
Samtools 0.1.17 0.1.12a
BowtieIndex (UCSC) (UCSC) (Ensemble)
.fa 2011.8.8 2011.8.6 2011.10.27 2011.8.6
.ebwt 2011.5.16 2011.5.16 2011.9.27 2011.6.13
Annotation gene.gtf 2011.5.17 2011.5.17 2011.9.27 2011.10.16
file ~/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/gene.gtf /GenomeDB/UCSC/Mus_musculus/mm9/Annotation/Genes/genes.gtf ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/gene.gtf /GenomeDB/MmGenome
cufflinks v1.0.3 v1.1.0 (2699)
cuffdiff v1.0.3 v1.1.0 (2699)

Data

  • make shortcut for input data:
ln -s /media/Ext6T/111005_SN853/RNAseq Hiseq111005/
  • make a shorcut for output data:
ln -s /media/Ext4T/DataDrive.backup/RL_Scratch/RNAseq RNAseq/
  • make copy of sequence file
cp Hiseq111005/* RNAseq/
  • combine different files in different lanes into one file for each index:
less *Indx3.txt > Indx3.txt
  • Transfer files to meangenemachine, as genome-miner is currently busy
ssh ruiliu@meangenemachine.dynamic.ucsd.edu
scp ruiliu@genome-miner:~/RNAseq/Hiseq111005/Indx_seq/* ./

Tophat_10.25.11 w/o G correction

Tophat_10.27.11 w/ G correction

Repeat RNAseq analysis on Hiseq111005 from 11.10.11


Tophat_11.10.11

  • Genome-minor from 11:15am on 11.10.11
  • Tophat 1.3.1
  • Re-do on Indx 10, 11, 13, 14, 15, and 3
  • move tophat folders into tophat_11.10.11
nohup tophat -p 3 --solexa1.3-quals -o Indx3 -G ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Sequence/BowtieIndex/genome ../Indx_seq/Indx3.txt &
nohup tophat -p 3 --solexa1.3-quals -o Indx10 -G ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Sequence/BowtieIndex/genome ../Indx_seq/Indx10.txt &
nohup tophat -p 3 --solexa1.3-quals -o Indx11 -G ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Sequence/BowtieIndex/genome ../Indx_seq/Indx11.txt &
nohup tophat -p 3 --solexa1.3-quals -o Indx13 -G ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Sequence/BowtieIndex/genome ../Indx_seq/Indx13.txt &
nohup tophat -p 3 --solexa1.3-quals -o Indx14 -G ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Sequence/BowtieIndex/genome ../Indx_seq/Indx14.txt &
nohup tophat -p 3 --solexa1.3-quals -o Indx15 -G ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Sequence/BowtieIndex/genome ../Indx_seq/Indx15.txt &

Cufflinks_11.11.11

Cuffdiff_11.11.11

  • UCSC gtf
nohup cuffdiff ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf -N -p 4 -o UCSC_cuffDiff_wtE13.5f_vs_KOE13.5f ../tophat_10.27.11/Indx6/Indx6_tophat/accepted_hits.bam,../tophat_11.10.11/Indx11/Indx11_tophat/accepted_hits.bam,../tophat_10.27.11/Indx12/Indx12_tophat/accepted_hits.bam ../tophat_11.10.11/Indx14/Indx14_tophat/accepted_hits.bam,../tophat_11.10.11/Indx15/Indx15_tophat/accepted_hits.bam &
nohup cuffdiff ~/RNAseq/iGenome/Mus_musculus/UCSC/mm9/Annotation/Genes/genes.gtf -N -p 4 -o UCSC_cuffDiff_wtE13.5m_vs_KOE13.5m ../tophat_10.27.11/Indx5/Indx5_tophat/accepted_hits.bam,../tophat_10.27.11/Indx9/Indx9_tophat/accepted_hits.bam,../tophat_11.10.11/Indx10/Indx10_tophat/accepted_hits.bam ../tophat_11.10.11/Indx13/Indx13_tophat/accepted_hits.bam &
  • Emsemble gtf
nohup cuffdiff /GenomeDB/MmGenome/Mus_musculus.NCBIM37.chr.64.gtf -N -p 8 -o Ensemble_cuffDiff_wtE13.5f_vs_KOE13.5f ../tophat_10.27.11/Indx6/Indx6_tophat/accepted_hits.bam,../tophat_11.10.11/Indx11/Indx11_tophat/accepted_hits.bam,../tophat_10.27.11/Indx12/Indx12_tophat/accepted_hits.bam ../tophat_11.10.11/Indx14/Indx14_tophat/accepted_hits.bam,../tophat_11.10.11/Indx15/Indx15_tophat/accepted_hits.bam &
nohup cuffdiff /GenomeDB/MmGenome/Mus_musculus.NCBIM37.chr.64.gtf -N -p 8 -o Ensemble_cuffDiff_wtE13.5m_vs_KOE13.5m ../tophat_10.27.11/Indx5/Indx5_tophat/accepted_hits.bam,../tophat_10.27.11/Indx9/Indx9_tophat/accepted_hits.bam,../tophat_11.10.11/Indx10/Indx10_tophat/accepted_hits.bam ../tophat_11.10.11/Indx13/Indx13_tophat/accepted_hits.bam &

Tophat against Ensemble on 11.14.11