1. Assembly of BAC ends on P250I21
Selection of BACs (IV)
Assembly of BAC ends on P250I21

2. Repetitive BAC end sequences on P308F14
Selection of BACs (V)
Repetitive BAC end sequences on P308F14
Perfect Blast score
High quality base discrepancy
Repetitive

3. AFLP fingerprints from chromosome 6 tomato BACs
Panel I, lanes 2 to 6 contains fingerprints for BACs from the Mi contig. Panel II, lanes 9 to 16 contains fingerprints for BACs from contig P103. All fingerprinted BACs originate from a HindIII BAC library except for lane 12 which was pulled from an MboI library. For all fingerprints EcoRI/MseI +0/+0 primer combinations have been used where +0 indicates the absence of selective nucleotides. Lane 1, 8, and 18 contain a 10 bp size marker. The molecular weight size range of the fingerprints is between 50 and 500 nts an indicated at the right size. BACs used for fingerprints are indicated below.
1 = 10_BL = P118F20
2 = P073H07 11= M054C01
3 = P096H22 12= P200L01
4 = P092A17 13= P061I06
5 = P111A08 14= P008K02
6 = P250I = P103N18
7 = P112G05 16= P188J12
8 = 10_BL = P197N20
9 = P046G10 18= 10-BL

4. Binning and high resolution physical map
Mi Contig
3,9 kb overlap

5. Results: 18 Seed BACs screened against 220.000 BAC ends
On average 7 overlapping BAC-ends per seed BAC
All ‘seeds’ have candidates from 2 libraries
M008H07 overlaps 3.6kb with P125P18
M027B07 overlaps 880bp with P153O03
M015K08 overlaps 5.0kb with P057J04
P103N18 overlaps 2.1kb with P046G10
P073H07 overlaps 3.9kb with P250I21
BlastN-analysis has been fully automated

6. Conclusions Proof of concept for the “BAC-end strategy”
Works for both fully closed BACs as for Phase 1 – 2 BACs
No need to fingerprint an entire BAC library
Blast, Assembly and AFLP work complementary
Accurate selection of BACs having minimal overlap and largest extending insert

7. Taco Jesse (Poster)
Kim Jansen
Dennis Woltinge
Jan van Haarst
Thamara Hesselink
Marjo van Staveren
Marleen Henkens
Paul Mooijman
Sander Peters (Poster)
René Klein Lankhorst