pre-genomic era: finding your own clones
DESCRIPTION
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. - PowerPoint PPT PresentationTRANSCRIPT
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Pre-genomic era: finding your own clones
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GENOME
CHROMOSOMES
YACsor
BACs (map)
PlasmidSub-clones
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Gridded (arrayed/ordered) library
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18,000 BACs for Drosophila
300,000 BACs for Humans
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STS- Sequence Tagged Site Mapping
STS Probe orPCR product
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Minimal Tiling Path
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Sequence 500-750ntFrom each end
Primer walk
PlasmidsOr M13
Sub-clone into smaller segments and mapOr use primer walking- NOT EFFICIENT
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Sub-clone
End sequences
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Virtual DNA(sequence)Assembly
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2 or 3 libraries ofDifferent size fragments
ShotgunSequencing
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Mates: read-pairs
2kb library of clones
10kb library of clones
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GENOME
CHROMOSOMES
YACsor
BACs (map)
PlasmidSub-clonesDon’t map Sequence 500-750nt
Assemble sequence of BAC
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Mates: read-pairs
2kb library of clones
10kb library of clones
150kb library of clones
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C. Elegans 100Mb
Drosophila 120Mb
Human 3,200Mb
Cosmids, YACs ordered
BACs ordered- STS mapping+ fingerprinting
BACs ordered- fingerprinting
Whole Genome shotgun
Whole Genome shotgun