Daniel:Notebook/RNAFISH/2014-9-3

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Trial Run (Started 08/29/2014)[edit]

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Post-Hybidization[edit]

  1. Remove rubber cement together with the chamber using a pair of fine forceps
  2. Wash slides in 2X SSC at 37°C, 3 × 15 min (preferably with shaking) to remove hybridization mixture
  3. Perform stringency washes in 0.1X SSC at 60°C, 2 × 5 min to remove nonspecific hybrids
  4. Equilibrate in 2X SSC for 2 min
  5. Add DAPI, let sit for at least 30 minutes
  6. Seal the coverslip with nail polish; let sit 30 minutes
    1. Sections become fragile after the FISH; therefore, strong pressure on the coverslip should be avoided
    2. e.g., pressing the coverslip with the microscope objective when focusing often completely destroys the sample

Microscopy[edit]

This sample was viewed over several days, they are listed here:

<dynamicpagelist> category=RNAFISH category=Microscopy category=20140829 </dynamicpagelist>