Hi everyone,

I generated DIG-labeled DNA probes via PCR for southern blot. In the DIG application manual for filter hybridization (roche), it is mentioned at p. 101 that these probes can be reused 3 - 5 times. I wonder if I have to heat the probe up again when I want to use it for the 2nd time?

When I use the probe for the first time, it is not yet in hybridization buffer and I denature the probe before I add it to the hyb. buffer. But when I store it at -20°C after the first use, it's already in hyb buffer. Does heating up the sample would harm the hyb buffer somehow? But on the other hand, if I do not heat it up, is the probe still denatured?

I am looking forward to your experience and thank you in advance!

Lena