Daniel:Notebook/HiResChrPaint/2014-1-24: Difference between revisions
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>Djacobse (Created page with "=Alu Control (Started 01/08/2014)= Back to Calendar ==Cot1 Labeling== Labeling lots of Cot1 DNA...") |
>Djacobse |
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Labeling lots of Cot1 DNA for FISH experiments. | Labeling lots of Cot1 DNA for FISH experiments. | ||
[[Daniel:Protocols/AlexaDyeCouple#ULYSIS|Following ULYSIS protocol]] | '''Samples''': | ||
*5 ug Cot1 DNA | |||
*The entire sample of [[Daniel:Notebook/HiResChrPaint/2014-1-13#Nanodrop Results]] | |||
*[[Daniel:Protocols/AlexaDyeCouple#ULYSIS|Following ULYSIS protocol]] | |||
1. Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C) | 1. Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C) | ||
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4. Add appropriate volume of dye to sample (see table 2); if necessary add labeling buffer to bring volume to 25 uL | 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 | 5. Incubate at 80C for 15 minutes; stop the reaction by plunging the tube into an ice bath | ||
5-1 | 5-1 Measure pre-column concentration using nanodrop | ||
6. Purify using a [[Daniel:Protocols/CentriSep|Centri-Sep column]] | 6. Purify using a [[Daniel:Protocols/CentriSep|Centri-Sep column]] |
Revision as of 22:37, 24 January 2014
Alu Control (Started 01/08/2014)
Cot1 Labeling
Labeling lots of Cot1 DNA for FISH experiments.
Samples:
- 5 ug Cot1 DNA
- The entire sample of Daniel:Notebook/HiResChrPaint/2014-1-13#Nanodrop Results
1. 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! 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 5-1 Measure pre-column concentration using nanodrop 6. Purify using a Centri-Sep column