Daniel:Notebook/RNAFISH/2014-9-3

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Revision as of 16:39, 24 September 2014 by >Djacobse (→‎Trial Run (Started 08/29/2014))
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Trial Run (Started 08/29/2014)

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Post-Hybidization

  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

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

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