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==Conclusion== *90sec pepsin digestion and very careful gentle washes helped keep much more of the tissue *37C incubation for 30min at the start of Ampligase reaction helps a LOT *20+hr RCA reaction led to larger rolonies, but since density isn't as high not an issue **Might be better for 20X 0.75NA objective *For decoding to validate with RNAscope (where tile image is 5.6mm wide starting from superficial edge) I will only be able to compare the 3.6mm after 2mm superficial layers **Check RNAscope quantification of genes to make sure there is changes in gene expression in those layers *[[Matt:LabNotes/2015-11-11 | Decoded here]]
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