Daniel:Notebook/HiResChrPaint/2013-7-25

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Revision as of 17:04, 25 July 2013 by >Djacobse (Created page with "=S2 Probe Prep (Started 7/23/2013)= Back to Main ==Dye Coupling== *Following [[D...")
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S2 Probe Prep (Started 7/23/2013)

Back to Main

Dye Coupling

0. Dry samples in a vaccuum centrifuge

For each

1. Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature
2. Resuspend DNA in 5 uL nfH20
3. Add 5 uL DNA and 3 uL of sodium bicarbonate buffer in PCR tube
4. Denature samples for 5 minutes at 95C, then snap cool using ice box
5. Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds
   *dye cannot be saved for later use.  Use immediately!
6. Add 2 uL dye to sample
7. Incubate in the dark for 1 hour
8. Add 10 uL nfH20 to sample
9. Purify using Centri Sep column
10. Measure concentrations using Microarray setting in nanodrop

Labeling Results