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Method: Prehybridization of Repeat-containing Probes with Total Human DNA

4/18/1990

C. Helms


Purpose:

Time required:

Procedure:

  1. Add 300 Ál unlabeled sonicated human placental DNA to the labeled probe ( ~500 Ál). If required for size marker detection, also add the appropriate amount of labeled lambda probe.
  2. Denature by heating 10 minutes in a boiling water bath, then add the probe+ unlabeled human DNA mix to 10-20 ml prehybridization mix (in a 50 ml orange-cap tube). Mix thoroughly by inverting the tube several times. Rock the tube at the incubation temperature (usually 50 degrees C) for 30 minutes to overnight.
  3. After the prehybridization period, add the probe + prehyb mix to the hybridization bag, and proceed with the hybridization as usual.

References: none