Pre-genomic era: finding your own clones

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Presentation transcript:

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

Minimal Tiling Path

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

Sub-clone End sequences

Virtual DNA (sequence) Assembly

Shotgun Sequencing 2 or 3 libraries of Different size fragments

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

* * * * * * * *

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

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

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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