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Tomato Genome Build
SL2.5 SL3.0
Surya Saha, Jeremy Edwards, Prashant Hosmani, Mirella Flores and
Lukas Mueller
Sol Genomics Network (SGN)
Boyce Thompson Institute, Ithaca, NY
ss2489@cornell.edu @SahaSurya
Slides: http://bit.ly/SOL15bld3
Tomato Genome
SGN Workshop, SOL 2015
Whole-genome shotgun and full-length
BAC sequences
• 454 reads (filtered, 22X)
• BAC end Sanger reads (0.18X)
• Fosmid end Sanger reads (0.087X)
• Selected BACs
• Sanger reads (5.2X).
Tomato Build SL2.40 SL2.50
SGN Workshop, SOL 2015
Lindsay Shearer
Stephen Stack
SL2.50 Availability
JBrowse
• Locus pages
• Gene pages
• FTP site
Also at NCBI
http://www.ncbi.nlm.nih.gov/assembly/GCF_000188115.3/
SGN Workshop, SOL 2015
CHROMOSOMES
SCAFFOLDSCONTIGS
Gene to Genome – The BIG picture
CONTIG GAPS
SCAFFOLD GAPS
SGN Workshop, SOL 2015
GENES
MAP (chr1)
Ovate (chr1)
TM2 (chr 9)
L2 (chr 10)
State of the SL2.50 Build
SGN Workshop, SOL 2015
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
0 1 2 3 4 5 6 7 8 9 10 11 12
Sequence Scaffold gap length Component gap length
State of the SL2.50 Build
SGN Workshop, SOL 2015
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
0 1 2 3 4 5 6 7 8 9 10 11 12
Sequence Scaffold gap length Component gap length
Length 823Mb
Sequence 737Mb
Contig gaps 43Mb (5.30%)
Scaffold gaps 42Mb (5.17%)
Total gaps 86Mb (10.47%)
Tomato BAC Resources
Bruce Roe
HTGS Phase 1: 332
HTGS Phase 2: 520
HTGS Phase 3: 2764
http://www.ncbi.nlm.nih.gov/genbank/htgs/faq
SGN Workshop, SOL 2015
ftp://ftp.solgenomics.net/tomato_genome/bacs/
SGN Workshop, SOL 2015
BAC libraries
• HindIII (129,024 clones)
• EcoRI (72,264 clones)
• MboI (52,992 clones)
Tomato BAC Resources…
2764 Phase 3 BACs
340,000 high-quality reads
20X of entire genome
Mueller et al. 2009
SGN Workshop, SOL 2015
ITAG 2.4 Genes
SL2.50 Genome
Whole Genome
Shotgun assembly
Phase 3 BACs
20kb or more
SGN Workshop, SOL 2015
ITAG 2.4 Genes
SL2.50 Genome
Whole Genome
Shotgun assembly
Phase 3 BACs
20kb or more
No BACs mapped
here!!
Workflow
SGN Workshop, SOL 2015
Automatic
integration of BACs
Manual validation NCBI validation
Workflow
SGN Workshop, SOL 2015
Automatic
integration of BACs
Manual validation NCBI validation
https://github.com/solgenomics/Bio-GenomeUpdate
BAC
assemblies
Align to SL2.50
• 500bp BAC ends
• 100% identity
Jeremy Edwards
Place
BACs
Phrap Assembly (HTGS Phase 3 BACs)
SGN Workshop, SOL 2015
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
1 2 3 4 5 6 7 8 9 10 11 12
Assembled BACs Singleton BACs
Phrap Assembly (HTGS Phase 3 BACs)
SGN Workshop, SOL 2015
Chr10 Contig68 10 BACs (242Kb!!)
Chr2 Contig185 7 BACs (566Kb!!)
Workflow
SGN Workshop, SOL 2015
Automatic
integration of BACs
Manual validation NCBI validation
Prashant Hosmani
Mirella Flores
Workflow
SGN Workshop, SOL 2015
Automatic
integration of BACs
Manual validation NCBI validation
SGN Workshop, SOL 2015
0
50
100
150
200
250
300
350
400
450
1 2 3 4 5 6 7 8 9 10 11 12
1041/2764 BACs Integrated
• No contained BACs
• No single-end mapping BACs
SGN Workshop, SOL 2015
Improved Contiguity
0
500
1000
1500
2000
2500
3000
1 2 3 4 5 6 7 8 9 10 11 12
SL2.50 components SL3.0 components
Gap Reduction
0
500000
1000000
1500000
2000000
2500000
3000000
3500000
4000000
4500000
1 2 3 4 5 6 7 8 9 10 11 12
SL2.50 Ns SL3.0 Ns
Future
BioNano optical maps
• Integrate sequences from chr 0 into chrs 1-12
• Validation
• Improved gap sizing
SGN Workshop, SOL 2015
Gabino Sanchez
Future…
Annotation of SL3.0
• Lift over ITAG 2.4 annotations from SL2.50
• Maker annotations
• ITAG 3.0 annotations
SGN Workshop, SOL 2015
Thank you!!
Questions??
SGN Workshop, SOL 2015
Slides: http://bit.ly/SOL15bld3

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Tomato Genome Build SL3.0 Improves Contiguity and Reduces Gaps

  • 1. Tomato Genome Build SL2.5 SL3.0 Surya Saha, Jeremy Edwards, Prashant Hosmani, Mirella Flores and Lukas Mueller Sol Genomics Network (SGN) Boyce Thompson Institute, Ithaca, NY ss2489@cornell.edu @SahaSurya Slides: http://bit.ly/SOL15bld3
  • 2. Tomato Genome SGN Workshop, SOL 2015 Whole-genome shotgun and full-length BAC sequences • 454 reads (filtered, 22X) • BAC end Sanger reads (0.18X) • Fosmid end Sanger reads (0.087X) • Selected BACs • Sanger reads (5.2X).
  • 3. Tomato Build SL2.40 SL2.50 SGN Workshop, SOL 2015 Lindsay Shearer Stephen Stack
  • 4. SL2.50 Availability JBrowse • Locus pages • Gene pages • FTP site Also at NCBI http://www.ncbi.nlm.nih.gov/assembly/GCF_000188115.3/ SGN Workshop, SOL 2015
  • 5. CHROMOSOMES SCAFFOLDSCONTIGS Gene to Genome – The BIG picture CONTIG GAPS SCAFFOLD GAPS SGN Workshop, SOL 2015 GENES MAP (chr1) Ovate (chr1) TM2 (chr 9) L2 (chr 10)
  • 6. State of the SL2.50 Build SGN Workshop, SOL 2015 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% 0 1 2 3 4 5 6 7 8 9 10 11 12 Sequence Scaffold gap length Component gap length
  • 7. State of the SL2.50 Build SGN Workshop, SOL 2015 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% 0 1 2 3 4 5 6 7 8 9 10 11 12 Sequence Scaffold gap length Component gap length Length 823Mb Sequence 737Mb Contig gaps 43Mb (5.30%) Scaffold gaps 42Mb (5.17%) Total gaps 86Mb (10.47%)
  • 8. Tomato BAC Resources Bruce Roe HTGS Phase 1: 332 HTGS Phase 2: 520 HTGS Phase 3: 2764 http://www.ncbi.nlm.nih.gov/genbank/htgs/faq SGN Workshop, SOL 2015 ftp://ftp.solgenomics.net/tomato_genome/bacs/
  • 9. SGN Workshop, SOL 2015 BAC libraries • HindIII (129,024 clones) • EcoRI (72,264 clones) • MboI (52,992 clones) Tomato BAC Resources… 2764 Phase 3 BACs 340,000 high-quality reads 20X of entire genome Mueller et al. 2009
  • 10. SGN Workshop, SOL 2015 ITAG 2.4 Genes SL2.50 Genome Whole Genome Shotgun assembly Phase 3 BACs 20kb or more
  • 11. SGN Workshop, SOL 2015 ITAG 2.4 Genes SL2.50 Genome Whole Genome Shotgun assembly Phase 3 BACs 20kb or more No BACs mapped here!!
  • 12. Workflow SGN Workshop, SOL 2015 Automatic integration of BACs Manual validation NCBI validation
  • 13. Workflow SGN Workshop, SOL 2015 Automatic integration of BACs Manual validation NCBI validation https://github.com/solgenomics/Bio-GenomeUpdate BAC assemblies Align to SL2.50 • 500bp BAC ends • 100% identity Jeremy Edwards Place BACs
  • 14. Phrap Assembly (HTGS Phase 3 BACs) SGN Workshop, SOL 2015 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% 1 2 3 4 5 6 7 8 9 10 11 12 Assembled BACs Singleton BACs
  • 15. Phrap Assembly (HTGS Phase 3 BACs) SGN Workshop, SOL 2015 Chr10 Contig68 10 BACs (242Kb!!) Chr2 Contig185 7 BACs (566Kb!!)
  • 16. Workflow SGN Workshop, SOL 2015 Automatic integration of BACs Manual validation NCBI validation Prashant Hosmani Mirella Flores
  • 17. Workflow SGN Workshop, SOL 2015 Automatic integration of BACs Manual validation NCBI validation
  • 18. SGN Workshop, SOL 2015 0 50 100 150 200 250 300 350 400 450 1 2 3 4 5 6 7 8 9 10 11 12 1041/2764 BACs Integrated • No contained BACs • No single-end mapping BACs
  • 19. SGN Workshop, SOL 2015 Improved Contiguity 0 500 1000 1500 2000 2500 3000 1 2 3 4 5 6 7 8 9 10 11 12 SL2.50 components SL3.0 components
  • 21. Future BioNano optical maps • Integrate sequences from chr 0 into chrs 1-12 • Validation • Improved gap sizing SGN Workshop, SOL 2015 Gabino Sanchez
  • 22. Future… Annotation of SL3.0 • Lift over ITAG 2.4 annotations from SL2.50 • Maker annotations • ITAG 3.0 annotations SGN Workshop, SOL 2015
  • 23. Thank you!! Questions?? SGN Workshop, SOL 2015 Slides: http://bit.ly/SOL15bld3