Daniel:Notebook/HiResChrPaint/2014-7-19: Difference between revisions
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>Djacobse (Created page with "=Dystrophin Probes (Started 7/18/2014)= Back to Calendar ==Dye Labeling== #Warm DMSO and bicar...") |
>Djacobse No edit summary |
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#Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature | #Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature | ||
#Add | #Add 5 uL DNA and 3 uL of sodium bicarbonate buffer | ||
#Denature samples for 5 minutes at 95C, then snap cool using ice box | #Denature samples for 5 minutes at 95C, then snap cool using ice box | ||
#Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds | #Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds | ||
##dye cannot be saved for later use. Use immediately! | ##dye cannot be saved for later use. Use immediately! | ||
#Add | #Add 2 uL dye to sample | ||
#Incubate in the dark for 1 hour | #Incubate in the dark for 1 hour | ||
#Add 5 uL 3M NaOAc and 40 uL nfH20 to sample | #Add 5 uL 3M NaOAc and 40 uL nfH20 to sample | ||
#Recommend column purification after incubation | #Recommend column purification after incubation |
Revision as of 20:15, 19 July 2014
Dystrophin Probes (Started 7/18/2014)
Dye Labeling
- Warm DMSO and bicarbonate labeling buffer and nuclease free water to room temperature
- Add 5 uL DNA and 3 uL of sodium bicarbonate buffer
- Denature samples for 5 minutes at 95C, then snap cool using ice box
- Dissolve 1 vial of reactive dye in 2 uL solvent; vortex ≥10 seconds
- dye cannot be saved for later use. Use immediately!
- Add 2 uL dye to sample
- Incubate in the dark for 1 hour
- Add 5 uL 3M NaOAc and 40 uL nfH20 to sample
- Recommend column purification after incubation