Daniel:Notebook/HiResChrPaint/2013-10-7
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V6S3 and V4S1 Amplification Amino Primers (Started 10/03/2013)
Dye Coupling
- V4S1-> Alexa 546
- V6S3-> Alexa 594
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