Daniel:Notebook/HiResChrPaint/2014-3-6: Difference between revisions
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>Djacobse (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)
Dye Labeling
- Following 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)
File:ULYSIS ReactionTable.png