Daniel:Notebook/HiResChrPaint/2014-1-24
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Alu Control (Started 01/08/2014)
Cot1 Labeling
Labeling lots of Cot1 DNA for FISH experiments.
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 Add 5 uL nfH2O, mix thoroughly, then extract 5 uL and put into 45 uL nfH2O (pre-column sample) 6. Purify using a Centri-Sep column