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Nuclear run-on

A method used to estimate the relative rate of transcription of a given gene, as opposed to the steady-state level of the mRNA transcript (which is influenced not just by transcription rates, but by the stability of the RNA). This technique is based on the assumption that a highly-transcribed gene should have more molecules of RNA Polymerase bound to it than will the same gene in a less-active state. If properly prepared, isolated nuclei will continue to transcribe genes and incorporate 32P into RNA, but only in those transcripts that were in progress at the time the nuclei were isolated. Once the Polymerase molecules complete the transcript they have in progress, they should not be able to re-initiate transcription. If that is true, then the amount of radiolabel incorporated into a specific type of mRNA is theoretically proportional to the number of RNA Polymerase complexes present on that gene at the time of isolation. A very difficult technique, rarely applied appropriately from what I understand.

Commonly used subcellular localization annotation analysis website WoLF PSORT

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