Daniel:Notebook/HiResChrPaint/2013-10-7

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V6S3 and V4S1 Amplification Amino Primers (Started 10/03/2013)

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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