Daniel:Notebook/HiResChrPaint/2013-5-22
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Probe Prep Test 2 (Started 5/10/2013)[edit]
Dye Coupling (Round 2)[edit]
After talking with Dr. Zhang yesterday, he suggested I try the dye coupling with just 1 reaction, and purify with a standard column purification.
Protocol[edit]
1. Warm DMSO, Sodium bicarbonate, and V4S3 in 37 degree shaker 2. Add 2.5 uL nf H20 to dna (should bring total to ~5) 3. Denature at 95C for 5 minutes 4. Add 3 uL sodium bicarb to reaction 5. Add 2 uL DMSO to dye, vortex ≥10 seconds 6. Add 2 uL dye/DMSO to reaction; vortex ≥10 seconds 7. Incubate in the dark for 1 hour 8. Add 40 uL nf H20 9. Column purification, add 10x PB, elute with 30 uL nfH20 10. Measure in nanodrop using microarray setting
Nanodrop Results[edit]
ng/uL DNA | pmol/uL dye | pmol/uL DNA | dye/probe | ug dye | Yield (%) |
16.5 | 2.2 | 0.62 | 3.564 | 0.495 | 11.0 |