DNA Methylation Sequencing Analysis
  • Introduction
  • Data Preparation
    • Locate the MethPipe Files
    • Download Utilities
    • Download Annotations
    • Annotation File Preparation – Defining Genomic Regions
  • Analysis Work Flow
    • DNA Methylation at Genomic Bins
    • DNA Methylation at CpG Islands
    • DNA Methylation at TFBS
    • DNA Methylation at Various Genic Structure Regions
    • DNA Methylation at Repeat Elements
    • Add CpG Islands Co-localization Information to HMR BED Files
    • Similarity and Differences of HMRs and PMDs from H1 and IMR90
      • HMRs
      • PMDs
  • Visualization Using R
    • Install R Libraries
    • Execute the R Scripts
  • An introduction of UCSC Genome Browser
    • General Usage
    • The Compressed Binary Index Format
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  1. Analysis Work Flow

DNA Methylation at TFBS

Same as Chapter 2.1, we used intersectBed and groupBy commands to calculate the average methylation values at each TF binding site.

# Console output

# intersectBed 
chr1    81184   81397   Rad21   1000    +       chr1    81320   81321   0
chr1    81184   81397   Rad21   1000    +       chr1    81334   81335   0
chr1    227464  227809  Rad21   602     +       chr1    227650  227651  0
chr1    227464  227809  Rad21   602     +       chr1    227656  227657  0
chr1    227464  227809  Rad21   602     +       chr1    227664  227665  0

# groupBy
chr1    81184   81397   Rad21   2       0
chr1    227464  227809  Rad21   5       0.1
chr1    227505  227800  CTCF    5       0.1
chr1    530491  530823  c-Myc   26      0.26923
chr1    535929  536126  Rad21   13      0.15385
cd ~/

bsub -q 16G -o stdout -e stderr "intersectBed -a Data/wgEncodeRegTfbsClustered.bed.gz -b /work3/NRPB1219/hg18_h1_meth.bedGraph -wa -wb | groupBy -i - -g 1-4 -c 10,10 -o count,mean | awk -F $'\t' 'BEGIN { OFS=FS } { print \$1,\$2,\$3,\$4,\$5,sprintf(\"%.4f\",\$6) }' > Output/wgEncodeRegTfbsClustered.h1.meth"

bsub -q 16G -o stdout -e stderr "intersectBed -a Data/wgEncodeRegTfbsClustered.bed.gz -b /work3/NRPB1219/hg18_imr90_meth.bedGraph -wa -wb | groupBy -i - -g 1-4 -c 10,10 -o count,mean | awk -F $'\t' 'BEGIN { OFS=FS } { print \$1,\$2,\$3,\$4,\$5,sprintf(\"%.4f\",\$6) }' > Output/wgEncodeRegTfbsClustered.imr90.meth"

Use bjobs to check the all jobs have completed and ls to check the files was in the "Output" folder.

ls -la ~/Output/wgEncodeRegTfbsClustered.*.meth

# Console output

-rw------- 1 s00yao00 s00yao00 15299532 2014-12-20 18:18 /home/s00yao00/Output/wgEncodeRegTfbsClustered.h1.meth
-rw------- 1 s00yao00 s00yao00 15299532 2014-12-20 18:18 /home/s00yao00/Output/wgEncodeRegTfbsClustered.imr90.meth
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Last updated 5 years ago

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