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Pre-genomic era: finding your own clones

Pre-genomic era: finding your own clones. GENOME. 1. 2. 3. 23. CHROMOSOMES. YACs. or. BACs (map). Plasmid Sub-clones. Gridded (arrayed/ordered) library. 18,000 BACs for Drosophila. 300,000 BACs for Humans. STS- Sequence Tagged Site Mapping. Probe or PCR product. STS.

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Pre-genomic era: finding your own clones

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  1. Pre-genomic era: finding your own clones

  2. GENOME 1 2 3 23 CHROMOSOMES YACs or BACs (map) Plasmid Sub-clones

  3. Gridded (arrayed/ordered) library

  4. 18,000 BACs for Drosophila 300,000 BACs for Humans

  5. STS- Sequence Tagged Site Mapping Probe or PCR product STS

  6. Minimal Tiling Path

  7. Sub-clone into smaller segments and map Or use primer walking- NOT EFFICIENT Plasmids Or M13 Sequence 500-750nt From each end Primer walk

  8. Sub-clone End sequences

  9. Virtual DNA (sequence) Assembly

  10. Shotgun Sequencing 2 or 3 libraries of Different size fragments

  11. Mates: read-pairs 2kb library of clones 10kb library of clones

  12. * * * * * * * *

  13. GENOME 1 2 3 23 CHROMOSOMES YACs or BACs (map) Plasmid Sub-clones Don’t map Sequence 500-750nt Assemble sequence of BAC

  14. Mates: read-pairs 2kb library of clones 10kb library of clones 150kb library of clones

  15. Cosmids, YACs ordered C. Elegans 100Mb Drosophila 120Mb BACs ordered- STS mapping + fingerprinting Whole Genome shotgun Human 3,200Mb BACs ordered- fingerprinting Whole Genome shotgun

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