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  Troubleshooting
      >> Distorted chromosome morphology
Possible Causes Suggested Solutions
          Specimen over-denatured
  • Reduce the slide denaturing time.
      >> High background
Possible Causes Suggested Solutions
          Slides are too dirty
  • Immerse the slides in 100% ethanol and wipe with clean paper
          Too much cell debris
  • Wash cell pellets with fresh fixative several times before dropping.
          Inadequate washing after hybridization
  • Remove the cover slip and wash again.
  • Check the pH and temperature of the wash solution.
  • Change washing solution.
      >> Weak or invisible signals
Possible Causes Suggested Solutions
          Probe expired or quenched
  • Check the expiration date of the probes.
  • Store the probes at -20 degree Celsius in the dark. 
          Air bubbles in the hybridization area
  • Clean the cover slips before use.
  • Apply cover slips carefully to avoid making bubbles.
          Probe solution dried out during hybridization
  • Make sure the cover slip is sealed by rubber cement completely.
  • Add sufficient water in the moist chamber.

 

          Inadequate denaturing of probes
  •  Make sure the porbes are denatured at 75 degree Celsius.
          Probes are too dilute
  • Use more concentrated probes.
          Inadequate denaturing of slides
  • Increase the denaturing temperature.
  • Increase the denaturing time.
  • Change denaturing solution.
          Incorrect filter used

 (i.e. TexasRed filter should not be used for TAMRA)

  • Select proper filters.
  • Select proper color of the probes for the filters.
          Exhausted filters
  •  Change the filters according to the product instructions.
          Exhausted mercury lamp
  • Change the mercury lamp after the lamp has been used for more than 200 hours.
 

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