Daniel:Notebook/HiResChrPaint/2014-8-21: Difference between revisions
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>Djacobse (Created page with "=Padlock Probes (Started 8/13/2014)= Back to Calendar ==Dye Labeling== Time for dye labeling ...") |
>Djacobse |
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==Dye Labeling== | ==Dye Labeling== | ||
Time for dye labeling of the padlock probes. | Time for dye labeling of the padlock probes. I'm going to label the padlock probes with Alexa 488 and I'm going to make a Cot1-546 for controls. | ||
#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 17:58, 21 August 2014
Padlock Probes (Started 8/13/2014)
Dye Labeling
Time for dye labeling of the padlock probes. I'm going to label the padlock probes with Alexa 488 and I'm going to make a Cot1-546 for controls.
- 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