Daniel:Notebook/HiResChrPaint/2014-8-12

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Revision as of 01:05, 13 August 2014 by >Djacobse (→‎Microscopy)
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FISH with DAPI (Started 08/10/2014)

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Hybridization

  1. Remove coverslip carefully and wash in pre-warmed 2X SSCT at 60C for 15 minutes
  2. Incubate in 2X SSCT at room temperature for 10 minutes
  3. Incubate in 0.2X SSC at room temperature for 10 minutes
  4. Dry with kimwipes; do not let the surface containing the cells touch the kimwipes
  5. Add 15 uL DRAQ5 (diluted 1:1000) to center of dish
  6. Cover with 22x22 #1.5 coverslip; seal with nail polish
    1. Allow at least 30 minutes for nail polish to dry
    2. Slides are quite stable for several months at 4C

Microscopy

Sample C

Sample A

Sample D <gallery> File:20140812_Cot1488DMD546.lif_Field1_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 1 (3.55X) File:20140812_Cot1488DMD546.lif_Field2_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 2 (3.55X) File:20140812_Cot1488DMD546.lif_Field2_1X_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 2 (1X) File:20140812_Cot1488DMD546.lif_Field3_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 3 (5X) <gallery>