Daniel:Notebook/HiResChrPaint/2013-5-20

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Probe Prep Test 2 (Started 5/10/2013)[edit]

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Dye Coupling[edit]

Protocol

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.