Hi,

I have a question regarding the detection of biotinilated probe in metaphase FISH, using old slides (made mid July 2014).

I use DIG~ and Biotin-Nick mixes to generate the probes. I try to detect biotinilated probe with avidin-FTIC. While I have almost no problem detecting DIG-labeled probes, I almost consistently fail to detect biotinilated probes.

Before hybridization I treat the slides with proteinase K (duration depends on the cell line) and do 2 min denaturation. I do an over night hybridization, usually around 14 h.

Thanks for your answers,

Jana