Why am I continuously getting positive signal in my 3 plex negative control in the RNAscope multiplex V2 kit?

To Ka Dorcas Cheung @To-Ka-Dorcas-Cheung

01 March 2024 0 5K Report

I am currently working to optimise RNAscope multiplex V2. I have troubleshooted the issue with several tests.

I have washed thoroughly with wash buffer even with their recommended guidelines of 200ml WB with slight agitation.

I have cleaned all of my containers with RNAase (even though not required, did it in case of probe contamination)

I wonder if this has anything to do with the TSA vivid dyes. Would it have a similar reaction to DAB where peroxidases could potentially create background signal? Would anyone recommend a stronger H2O2 quench?

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