Genomiphi

The 51 loci were validated in a combined European sample of 3,893 cases and 2,829 diverticula-free controls based on colonoscopy (Table 1) using the most significant discovery variant. When direct genotyping of a lead variant was not technical feasible, appropriate proxies were selected instead, defined as the variant with the next-lowest P-value within 250 kb of the index SNP (Table 2,3 Supplementary Table 5 ). Genotyping of SNPs was performed using the Agena® iPLEX Gold chemistry MassARRAY platform and TaqMan® technology from Life Technologies on an automated platform as described previously [2] . The choice of genotyping technology per variant was based on technical considerations of assay design feasibility and is indicated in Table 2 and3. Genomic DNA was amplified with the GenomiPhi (Amersham) whole-genome amplification kit and fragmented at 99 °C for 3 min. All data were logged and managed with a database-driven laboratory information management system (LIMS) [7] . Individual samples with >5% missing data were excluded from

DNA was prepared using the FlexiGene chemistry (Qiagen, Hilden, Germany). For preparation of genotyping, DNA samples were evaluated by gel electrophoresis and adjusted to 20-30 ng/µl DNA content using the Picogreen fluorescent dye (Molecular Probes -Invitrogen, Carlsbad, Ca, USA) on a robotic platform using TECAN liquid handling equipment. One microliter of genomic DNA was amplified with the GenomiPhi (Amersham, Uppsala, Sweden) whole genome amplification kit and fragmented at 99°C for three minutes. Five ng of DNA was performed using TaqMan® SNP Genotyping Assays and chemistries (Applied Biosystems, Foster City, CA, USA) on an automated platform with TECAN Freedom EVO and 384well TEMO liquid handling robots (TECAN, Männedorf, Switzerland) as described before. [12] All process data were logged and administered with a database-driven LIMS. Reactions were completed and read in a 7900 HT TaqMan sequence detector system (Applied Biosystems, Foster City, CA, USA). The amplification reaction was carried out with the TaqMan universal master mix. Thermal cycling conditions consisted of 1 cycle for 10 minutes at 95°C, followed by 45 cycles for 15 seconds at 95°C, and 45 cycles for 1 minute at 60°C.