Daniel:Notebook/HiResChrPaint/2013-11-21
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Positive Control 2 Started 11/14/2013)
ssNanodrop Results
Following is the single stranded nanodrop results from my 4 probe preps so far.
Sample | ng/uL | ug in 20 uL |
V6S1 | 103.7 | 2.1 |
V6S2 | 226.9 | 4.5 |
V6S3 | 144.3 | 2.9 |
V6S4 | 137.2 | 2.7 |
Dye Coupling
This will be the same as usual dye couplings, except that I'm going to label all dyes with Alexa 488, and I'm going to pool two of the samples together (V4S1,V4S2 and V4S3,V4S4) together before dye coupling, since the labeling is supposed to be for 1-5 ug of DNA, and each of these only has around 2 ug.
Protocol
For each, from ARES Protocol
0. Dry each sample in vacuum centrifuge, resuspend in 5 uL nfH20 1. Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature 2. Add 5 uL DNA and 3 uL of sodium bicarbonate buffer 3. Denature samples for 5 minutes at 95C, then snap cool using ice box 4. Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds *dye cannot be saved for later use. Use immediately! 5. Add 2 uL dye to sample 6. Incubate in the dark for 1 hour 7. Add 15 uL nfH20 to sample 8. Centri Sep column purification