Daniel:Notebook/HiResChrPaint/2013-8-10
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S2 Probe Prep (ULS; Started 8/8/2013)[edit]
Dye Coupling[edit]
I have 2 uL left of the dye, so I can only couple V4S2, which has ~2 ug left (you need 1 uL dye/ug DNA).
0. Dry sample in vaccuum centrifuge
- Following ULYSIS Protocol
1. Resuspend DNA in 20 uL labeling buffer (Component C) 2. Denature DNA for 5 minutes at 95C; snap cool on ice 3. Add 1 uL dye to sample per ug 4. Incubate for 20 minutes at 80C, snap cool on ice 5. Purify using Centri Sep column
Labeling Results[edit]
ng/uL DNA | pmol/uL dye | ug DNA in 25 uL | pmol/uL DNA | dye/probe | dye/100 bp | Base:Dye Ratio (:1) | |
V4S2 | 85.1 | 2 | 2.1 | 3.2 | 0.6 | 0.8 | 129 |