hybridization with heterologous probe

Van Geldre Els Els.VanGeldre at rug.ac.be
Thu Jun 12 19:26:54 EST 1997


Hi netters,

Does anyone of you specialists know something about hybridization 
conditions with heterologous probes ? I'd like to perform the 
hybridization in Dig Easy Hyb. Boehringher Mannheim gives stringent 
conditions if probe and target have 100 % homology (temperature between 
37-42 °C). I will use a heterologous probe in order to identify any 
"specific PCR product" (the same enzyme as my probe, yet from another 
plant) obtained with degenerate primers (based on homologous boxes found 
after alignation of already isolated enzymes) on gDNA from my plants. 
Other experiments to obtain a "specific" PCR product with these primers 
failed (playing with annealing temperature, MgCl2 concentrations, nested 
PCR). Maybe someone has even a better suggestion than "Southern blot" on 
my PCR products followed by hybridization with a heterologous probe ?
Thank you for your attention,
Els

My E-mail : Els.VanGeldre at rug.ac.be



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