Daniel:Notebook/HiResChrPaint/2013-5-20
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Probe Prep Test 2 (Started 5/10/2013)[edit]
Dye Coupling[edit]
1. Warm reagents (bicarb,h20,DMSO) to 37C 2. Add 2.5 uL DNA and 1.5 uL nfH20 to 0.2 mL tube 3. Denature at 95C for 5 minutes and snap cool using ice box 4. Add 2.2 uL DMSO to dye, vortex ≥10 seconds 5. Add 1 uL dye to each reaction 6. Incubate in the dark for 1 hour 7. Add 5 uL 3M NaOAc and 40 uL nfH20 8. Purify using ssDNA column 9. Check using nanodrop; use microarray setting
Nanodrop Results[edit]
Sample | Starting ng/uL | Starting ug | ng/uL | ug total | Yield | pmol/uL (Alexa-594) |
V4S3 | 1798.9 | 4.5 | 190 | 1.9 | 42.2 | 0 |
V6S3 | 1510.1 | 3.8 | 192.1 | 1.921 | 50.9 | 0 |
Cells[edit]
Need to get some cells (fibroblasts, I think) from Alan. He is checking them today but they might not be ready until Wednesday. If they are ready today I will be fixing them and getting ready for hybridization.