FISH probe: random prime or nick translate?

Alberto Catalano acatalan at mail.usyd.edu.au
Fri Mar 28 02:20:30 EST 1997


W R Bennett wrote:

  Are there any good reasons not to use a random priming method to
  generate
  biotinylated or dig-labelled probes for FISH?  The protocols I have
  all
  use nick translation, but - at least in Southerns - we've had a lot
  more
  success with random primed probes than with nick translation.

  Thanks,

  Bill Bennett
  W.R.BENNETT at bath.ac.uk

 Don't forget that random prime labelling leaves some unlabelled
template. Depending on the reaction conditions you may end up with more
unlabelled DNA than labelled. This may be a problem when doing
chromosome in situ hybridizations on single copy sequences.

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|  Alberto Catalano                                                   |

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