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11/6/2014 graphSVG.svg
Background 
• “Pan Genome” - way to think about, compute 
on, visualize the differences and similarities of 
many genomes at once 
• Reference free structure 
• Many, many genomes
de Bruijn Graph Construction 
• Dk = (V,E) 
• V = All length-k subfragments 
• E = Directed edges between consecutive subfragments 
• Nodes overlap by k-1 words 
It was the best of It was the best was the best of 
• Locally constructed graph reveals the global sequence structure 
• Overlaps between sequences implicitly computed 
Slide: http://cbcb.umd.edu/confcour/CMSC828H-materials/ 
Lecture12-MSchatz-DeBruijnAssembly.pptx 
Original Fragment Directed Edge 
de Bruijn, 1946 
Idury and Waterman, 1995 
Pevzner, Tang, Waterman, 2001
Strategy: find all k-mers, build graph 
• Every k-mer becomes a node 
• Two nodes are linked with an edge if they 
share a k-1 mer 
GACTGGGACTCC 
GACTGG ACTGGG 
CTGGGA TGGGAC 
GGACTC GGGACT 
GACTCC
Strategy: k-mers from feature families, 
build graph 
• Every k-mer becomes a node 
– If it is present in m genomes 
• Two nodes are linked with an edge if they 
share a k-1 mer 
• d# = a feature family 
d1d2d3d4d5d6d7d8 
d9 
d1d2d3d4d5d 
6 
d1d2d3d4d5d6d7d8 
d2d3d4d5d6d 
7 
d3d4d5d6d7d8 d4d5d6d7d8d9 
d9
rf-graph de Bruijn “like”
Create pg-graph
Similarities and Differences 
10 groups of 10 
Organism Sum Pairwise Distances (Phylogenetic) 
E. coli 0.07 
Coxiella 10.42 
Mycobacterium 2.70 
Brucella 0.08 
Rickettsia 8.62 
Burkholderia 7.21 
Clostridium 9.05 
Bacillus 4.48 
Staph. 2.08 
Strep. 4.79
Similarities and Differences 
0.9 
0.8 
0.7 
0.6 
0.5 
0.4 
0.3 
0.2 
0.1 
0 
Node Increase vs. MUMi 
0.850 0.900 0.950 1.000 
Node Increase 
MUMi 
Node Increase vs. Diversity Score 
0.9 
0.8 
0.7 
0.6 
0.5 
0.4 
0.3 
0.2 
0.1 
0 
0.00 2.00 4.00 6.00 8.00 10.00 12.00 
Node Increase 
Diversity Score 
Node Increase = (Nodes – Max(Families)) / Nodes 
Diversity Score= Sum of maximum pairwise distances in Order level tree 
MUMi= Maximum of all pairwise MUMi in a group
Layout 
Gephi Toolkit 
Yifan Hu’s Multilevel 
Force Atlas 2
Colors and Lines
Dealing with many Genomes 
N=2 
K=5 
M=2 
B. Abortus 
N=40, K=5, M=2, B. Suis 
N=20 
K=5 
M=2 
Brucella 
N=400, K=5, M=2, All Brucella N=1000, K=10, M=100, E. coli
Information Compounded
For the Web 
• GEXF 
– NetworkX, Gephi, 
– Cytoscape, Gexf-JS, D3-Gexf 
• BGZF GFF 
– Backing store 
– Byte range loading
Other Uses 
• “Rearrangement” detection
Other Uses 
• “Scaffolding” 
– e.g. 86 contigs 
• Closing 
– Predicted primers
Other Uses 
• Rearrangements 
– Insertions/Deletions 
– Islands 
– Inversions
Other Uses 
• Synthetic BAM
Takeaways 
• A new way to leverage protein family 
databases 
• “Reference free” structure for many bacterial 
genomes using feature families 
• Quickly investigate whole genome 
relationships and speed up potentially 
expensive calculations
Acknowledgements 
• Eric Nordberg 
• Lenny Heath 
• CID at VBI (PATRIC) 
• RAST – Argonne (PATRIC) 
https://github.com/aswarren 
https://twitter.com/aswarren

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Pan-genome Graphs biodata14

  • 1.
  • 3. Background • “Pan Genome” - way to think about, compute on, visualize the differences and similarities of many genomes at once • Reference free structure • Many, many genomes
  • 4. de Bruijn Graph Construction • Dk = (V,E) • V = All length-k subfragments • E = Directed edges between consecutive subfragments • Nodes overlap by k-1 words It was the best of It was the best was the best of • Locally constructed graph reveals the global sequence structure • Overlaps between sequences implicitly computed Slide: http://cbcb.umd.edu/confcour/CMSC828H-materials/ Lecture12-MSchatz-DeBruijnAssembly.pptx Original Fragment Directed Edge de Bruijn, 1946 Idury and Waterman, 1995 Pevzner, Tang, Waterman, 2001
  • 5. Strategy: find all k-mers, build graph • Every k-mer becomes a node • Two nodes are linked with an edge if they share a k-1 mer GACTGGGACTCC GACTGG ACTGGG CTGGGA TGGGAC GGACTC GGGACT GACTCC
  • 6. Strategy: k-mers from feature families, build graph • Every k-mer becomes a node – If it is present in m genomes • Two nodes are linked with an edge if they share a k-1 mer • d# = a feature family d1d2d3d4d5d6d7d8 d9 d1d2d3d4d5d 6 d1d2d3d4d5d6d7d8 d2d3d4d5d6d 7 d3d4d5d6d7d8 d4d5d6d7d8d9 d9
  • 7. rf-graph de Bruijn “like”
  • 9. Similarities and Differences 10 groups of 10 Organism Sum Pairwise Distances (Phylogenetic) E. coli 0.07 Coxiella 10.42 Mycobacterium 2.70 Brucella 0.08 Rickettsia 8.62 Burkholderia 7.21 Clostridium 9.05 Bacillus 4.48 Staph. 2.08 Strep. 4.79
  • 10. Similarities and Differences 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 Node Increase vs. MUMi 0.850 0.900 0.950 1.000 Node Increase MUMi Node Increase vs. Diversity Score 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 0.00 2.00 4.00 6.00 8.00 10.00 12.00 Node Increase Diversity Score Node Increase = (Nodes – Max(Families)) / Nodes Diversity Score= Sum of maximum pairwise distances in Order level tree MUMi= Maximum of all pairwise MUMi in a group
  • 11. Layout Gephi Toolkit Yifan Hu’s Multilevel Force Atlas 2
  • 13. Dealing with many Genomes N=2 K=5 M=2 B. Abortus N=40, K=5, M=2, B. Suis N=20 K=5 M=2 Brucella N=400, K=5, M=2, All Brucella N=1000, K=10, M=100, E. coli
  • 15. For the Web • GEXF – NetworkX, Gephi, – Cytoscape, Gexf-JS, D3-Gexf • BGZF GFF – Backing store – Byte range loading
  • 16. Other Uses • “Rearrangement” detection
  • 17. Other Uses • “Scaffolding” – e.g. 86 contigs • Closing – Predicted primers
  • 18. Other Uses • Rearrangements – Insertions/Deletions – Islands – Inversions
  • 19. Other Uses • Synthetic BAM
  • 20. Takeaways • A new way to leverage protein family databases • “Reference free” structure for many bacterial genomes using feature families • Quickly investigate whole genome relationships and speed up potentially expensive calculations
  • 21. Acknowledgements • Eric Nordberg • Lenny Heath • CID at VBI (PATRIC) • RAST – Argonne (PATRIC) https://github.com/aswarren https://twitter.com/aswarren