1. Saliva sample collection with using Oragene®DNA kit DNA extraction & purification Gel electrophoresis Experimental Outline DNA quality and quantity assessment Sequenom® MassARRAY Genotyping 3730xL DNA Analyzer Sanger Sequencing 1 NanoDrop OD

2. 1.0% Agarose Gel Electrophoresis 10,000bp 4,000bp 2,000bp 800bp 400bp W1W3D1D2 50ng DNA /lane W4D3D4 Sample IDng/µlA 260 A 280 A 320 Estimated yield from 500ul sample (µg)(A 260 -A 320 ) / (A 280 -A 320 ) D1405.758.1154.4690.2540.5751.86 D2416.018.324.6170.27941.6011.85 W111.530.2310.1530.0331.1531.65 D3432.28.6444.8540.29843.221.83 D4434.178.6834.8570.29343.4171.84 W312.420.2480.1710.0131.2421.49 W415.330.3070.2090.0151.5331.51 2

3. Quality & Yield 3 Corrected A 260 /A 280 Estimated yield From 500uL Saliva (µg)

4. Downstream Applications Sequenom® MassARRAY SNP Genotyping 3730xl DNA Analyzer Sanger Sequencing

5. Sanger DNA Sequencing Results 5

6. Sequenom® MassARRAY Results 6 Percentage call rate (out of total 29 SNPs genotyped) Sample WSample D

7. Our Experience 7 13 Extractions done 7 Individuals Easy to use Hands-on time = 1.5-2.5 hrs Good quality With individual variations Data successfully generated for: Sanger DNA sequencing Sequenom MassArray Genotyping

8. Acknowledgements  Lauren Scully  All GRC colleagues 8