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Saliva sample collection with using Oragene®DNA kit DNA extraction & purification Gel electrophoresis Experimental Outline DNA quality and quantity assessment.

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Presentation on theme: "Saliva sample collection with using Oragene®DNA kit DNA extraction & purification Gel electrophoresis Experimental Outline DNA quality and quantity assessment."— Presentation transcript:

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 ) D D W D D W W

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 = hrs Good quality With individual variations Data successfully generated for: Sanger DNA sequencing Sequenom MassArray Genotyping

8 Acknowledgements  Lauren Scully  All GRC colleagues 8


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