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===Validation of results=== *f1 0.45 average abs; 5.202 uM, 3.0 mL total, final of 1.2 mL, 42.86% concentration, '''7.43 uM''' *f2 0.64 average abs; 7.398 uM, 3.4 mL total, final of 1.4 mL, 41.66% concentration, '''10.48 uM''' *calculation for uM. 100 uM/8.65 = X uM/abs '''10/28/2014''' *'''incubation with transposase only to test nuclease contamination''' *with 6 ng DNA *incubated reaction at 55C for 5 mins, followed by qiagen protease digestion, 50C for 10 mins, 70C for 20 mins. *reaction composition 1 uL pure Jurkat DNA (6 ng) 2-3 uL H2O 1-2 uL transposase (diluted if stated) __________ 5 uL total samples. 1. R7f1, 2 uL tsase, 6 ng DNA 2. R7f1, undil. tsase, 6 ng DNA 3. R7f1, 2X dil. tsase, 6 ng DNA 4. R7f1, 4X dil. tsase, 6 ng DNA 5. R7f1, 8X dil. tsase, 6 ng DNA 6. R7f2, 2 uL tsase, 6 ng DNA 7. R7f2, undil. tsase, 6 ng DNA 8. R7f2, 2X dil. tsase, 6 ng DNA 9. R7f2, 4X dil. tsase, 6 ng DNA 10. R7f2, 8X dil. tsasa, 6 ng DNA 11. nextera, 6 ng pure DNA *still has nuclease activity. given washing conditions, would lean more toward tn5 having a inherent nuclease activity. [[File:ZhangLab 2 2014-10-28 12hr 32min-labeled.jpg|600px]]
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