Rnasezap solution

Procedure was described previously in Földy et al., 2016 (link). To minimize interference with subsequent molecular experiments, only a small amount of intracellular solution (~1 μl; not autoclaved or treated with RNase inhibitor) was used in the glass pipette during electrophysiological recordings. Before and during recording, all surface areas - including manipulators, microscope knobs, computer keyboard, etc. - that the experimenter needed to contact during the experiment were cleaned with RNaseZAP solution (Sigma). After whole-cell patch-clamp recordings, the cell’s cytosol was aspirated via the glass pipette used during the recording. Although the aspirated cytosol may have contained genomic DNA, our choice of cDNA preparation, which involved poly-A based mRNA selection, virtually eliminated the possibility of genomic contamination in the RNAseq data. For sample collection, we quickly removed the pipette holder from the amplifier head stage and used positive pressure to expel samples into microtubes containing cell collection buffer while gently breaking the glass pipette tip. Cell collection microtubes were stored on ice until they were used.