Daniel:Notebook/HiResChrPaint/2014-7-19: Difference between revisions

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(Created page with "=Dystrophin Probes (Started 7/18/2014)= Back to Calendar ==Dye Labeling== #Warm DMSO and bicar...")
 
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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 2.5 uL DNA and 1.5 uL of sodium bicarbonate buffer
#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 1 uL dye to sample
#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)

Back to Calendar

Dye Labeling

  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
    1. 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 5 uL 3M NaOAc and 40 uL nfH20 to sample
  8. Recommend column purification after incubation