In order to validate mRNA expression in the mouse brain following RNAseq, I would like to do quantitative ISH. If you have any recommendations of which technique to use or company suggestions, where to acquire which probe, I would be very grateful.
Gary Steven McDowell
ISH to verify expression seems fine but quantitative ISH is very unreliable; the safer bet might be qPCR.
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Gary Steven McDowell
Right absolutely - but my only concern is the use of ISH as a robust quantitative measure. As you point out above, it's tricky to control for, and to image well for quantitative analysis. I guess my point would be that if the quantitative information already exists from the RNASeq, why try to validate it quantitatively using a poorer measure, rather than just image where it is expressed?
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