Daniel:Notebook/HiResChrPaint/2013-8-10

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S2 Probe Prep (ULS; Started 8/8/2013)[edit]

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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
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