1. Sequence Capture and Targeted Re-sequencing
Thahira Rahman, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne

2. Sequence Capture / Genome reduction:
Ultra deep sequencing

3. Modifications done to Agilent’s in-solution hybrid capture method
Sample QC_ 260/230 and 260/280: 2.0 to 2.2
Fragmentation on Covaris

4. Covaris fragment profiles observed on DNA 1000 LabChip

8. End Repair
Klenow exo- and ATP

9. Ligate Multiplex PE adapters
SPRI bead purification
Ligated samples checked on DNA 1000 LabChip (please refer previous slides)

10. Custom synthesised Blocks used for Hyb
Probes designed on eArray
Adapter linked gDNA fragments
SureSelect Biotinylated RNA Baits
Hybridisation 65C for 24h)
Streptavidin coated magnetic beads
Hybrid capture using MPC
Unbound fragments
Custom synthesised Blocks used for Hyb

11. Post hybridisation PCR involve indexing of libraries
Quantification of adapted and enriched libraries on High Sensitivity LabChip

12. Equimolar pooling of libraries
50bp Multiplex PE Illumina sequencing
Data Analyses

13. The data was aligned with hg18 genome build using bowtie
Size of the target region covered by SureSelect Human X-Chromosome baits is
3,053,381 bp.
Total length of the captured sequence is 3,025,546 bp (99.09%).

15. Coverage achieved:
Maximum: x
Average: x
Minimum: 50.95x