physical maps of mouse, medaka, sugar beet …. and ... · pooled 35mer oligo probes on...

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Heinz Himmelbauer Max-Planck-Institute of Molecular Genetics Berlin, Germany Physical maps of mouse, medaka, sugar beet …. and Physcomitrella

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Page 1: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Heinz HimmelbauerMax-Planck-Institute of

Molecular GeneticsBerlin, Germany

Physical maps of mouse, medaka, sugar beet …. and Physcomitrella

Page 2: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Construction of a physical YAC/BAC map

of the mouse genome

IRS-PCR principle and application

Probe A Probe B

Strain X

Strain Y

IRS-products fromYAC pool DNA

IRS-products fromRH line DNA

IRS-products fromgenetic cross DNA

YAC clones Position onRH map

Position ongenetic map

YAC pool filter hybridisation

Scoring tool for data entry

Page 3: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Advanced mouse YAC map including BAC frameworkdata access at www.molgen.mpg.de/~rodent

- Browse the map- Identify map intervals and clones- Obtain clones and markers- Query database- Download data

Page 4: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Phylogenetic position of the medaka

Page 5: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Probe poolplate1/A5-H5

Probe poolplate1/B1-B6

Pooled 35mer oligo probes on 36,864-clone medaka BAC array

Page 6: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Total number of medaka BAC map segments (mapped, ambiguous, unmapped)1st generation map (January 2004)

464 4299

758

88

1878

0

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1000

1200

1400

1600

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2000

M A Un

ProbesMap segments

Page 7: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Sugar beet (Beta vulgaris) BAC mapProject launched Sept 1st, 2004

Goal:

Phase-1 map:

* 10,000 probes (ESTs, BAC ends, genetically mapped markers) as 35-mer oligos* mapped onto 37,000 BACs (120 kb avg. insert size / 5 x coverage)* 1000 contigs, 800 kb avg. length

Phase-2 map* Further 2000 probes from contig ends* additional Bac resources (avg. insert size 160 kb)* final product: 500 contigs, 1.6 Mb avg. length

Page 8: Physical maps of mouse, medaka, sugar beet …. and ... · Pooled 35mer oligo probes on 36,864-clone medaka BAC array . Total number of medaka BAC map segments (mapped, ambiguous,

Proposal for a Physcomitrella BAC map

Assumption: BAC library (36,000 clones avg. insert size 120 kb, 8.4 x coverage) available

Phase-1 map:

* 6000 BAC end probes (3000 BAC clones) as 35-mer oligos* 1000 ESTs* ~ 700 contigs, ~700 kb avg. length* ~ ~1500 genetically mapped markers are needed

Phase-2 map* Further 1400 probes from contig ends* final product: ~350 contigs, ~1.4 Mb avg. length