File(s) under embargo
until file(s) become available
We designed FRET-FISH probes for optimization of the design with the first being on MYC gene on human HAP1 cell lines and Ogt on mouse NIH3T3 and MEF.
Thereafter, the optimized FRET-FISH design targeted Magix, Kdm5c,
Atp2b3, Tent5d, Ddx3x and Pbd1c. A FRET-FISH probe spans around 100 kb. The
experiments were replicated.
Experiment Imaging Method: epifluorescence microscopy
Protocol Name: FRET-FISH protocol
Each dataset consists of multiple fields of view (FOVs) that were acquired with 2 different dyes and the FRET channel that consists using the excitation source of the donor dye and filter the fluorescence signal with the emission filters of the acceptor dye. (AF488 labeled as ‘a488’; Cy3 as ‘tmr’; AF594 as ‘a594’; Cy5 as ‘Cy5’ and Hoechst 33342 as ‘dapi’) and all the FOVs are provided as separate .tiff files. Each FOV comprises 40–50 focal planes spaced 0.3 μm apart and has a size of 1024x1024 pixels.