Daniel:Notebook/HiResChrPaint/2014-3-6: Difference between revisions

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(Created page with "=Overload FISH (Started 3/5/2014)= Back to Calendar ==Dye Labeling== *Following [[Daniel:Protoc...")
 
>Djacobse
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*Following [[Daniel:Protocols/AlexaDyeCouple#ULYSIS|ULYSIS protocol]]
*Following [[Daniel:Protocols/AlexaDyeCouple#ULYSIS|ULYSIS protocol]]
#Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C)
##For AlexaFluor 488 label using 5 uL DMSO (stable for 1 month at 4C)
##VORTEX THESE SOLUTIONS '''VIGOROUSLY''' BEFORE USE '''EVERY TIME'''!
#Resuspend DNA in 20 uL labeling buffer (Component C)
#Denature DNA at 95C for 5 minutes and snap cool on ice; centrifuge briefly
#Add appropriate volume of dye to sample (see table 2); if necessary add labeling buffer to bring volume to 25 uL
#Incubate at 80C for 15 minutes; stop the reaction by plunging the tube into an ice bath
#Purify using a column (Centri-Sep recommended)
[[Image:ULYSIS_ReactionTable.png|500px]]

Revision as of 17:16, 6 March 2014

Overload FISH (Started 3/5/2014)

Back to Calendar

Dye Labeling

  1. Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C)
    1. For AlexaFluor 488 label using 5 uL DMSO (stable for 1 month at 4C)
    2. VORTEX THESE SOLUTIONS VIGOROUSLY BEFORE USE EVERY TIME!
  2. Resuspend DNA in 20 uL labeling buffer (Component C)
  3. Denature DNA at 95C for 5 minutes and snap cool on ice; centrifuge briefly
  4. Add appropriate volume of dye to sample (see table 2); if necessary add labeling buffer to bring volume to 25 uL
  5. Incubate at 80C for 15 minutes; stop the reaction by plunging the tube into an ice bath
  6. Purify using a column (Centri-Sep recommended)
File:ULYSIS ReactionTable.png