Daniel:Notebook/HiResChrPaint/2014-7-15: Difference between revisions

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==Dye Labeling==
==Dye Labeling==
I need to order more ULYSIS 488, so for now I'll just do 2 ug of V6S2 labeled with Alexa 546.


#Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C)
#Add 100 uL DMSO to vial containing ULS dye (stable for 6 months at 4C)

Revision as of 21:23, 15 July 2014

Strand Control (Started 07/08/2014)

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Dye Labeling

I need to order more ULYSIS 488, so for now I'll just do 2 ug of V6S2 labeled with Alexa 546.

  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)