DNA probes for In Situ Hybridization - equivalence of Sense probe for control for Dig-DNA-label probe? (Dec/15/2010 )
I am trying to design my ISH experiment. I had already DNA-DIG label probe which work pretty well for my Northern-blot and I want to use this same probe for my ISH.But, in most of the papers I readed, the ISH are performed with RNA probe and the sense probe is used as control.
My problem is that I really don't know what can I used as control for my ISH performed with DNA-DIG label probe.
Someone have an idea?
If possible I would like to skip the solution of doing a construction with a T3 or T7 promoter, because of the time it would take me)
Thanks for the help,
You can use DNA, the problem is that DNA:RNA hybrids are not as stable as RNA:RNA - so there is slightly more chance of degradation. For a control you could use the complement of your probe - you would have to make this though (synthesise as you would for long primers?).