We employed individual inspection of loci in order to visualize differences between replicates and to confirm the correct identification of bound regions QC).
We used TCS v1.21 [ 43] to generate haplotype networks for both loci in order to visualize the geographic distribution of haplotypes, and to determine if the data fit the assumption of being "tree-like".
We tagged a nuclear pore protein, Nup49, in its chromosomal locus with GFP to visualize the nuclear membrane and used confocal microscopy to capture three-dimensional images of the nucleus to quantify volumes of ≥150 cells per H2A mutant.
The Scroll/Zoom tools allow the user to visualize loci throughout the entire genome at variable resolution levels.
To visualize loci near ARS727 and ARS731, tetOx224 (11.2 kb) and lacOx256 (10.1 kb) were integrated by a two-step "pop-in and pop-out" method.
Moreover, FISH and immunocytochemical techniques can underestimate the number of genetic loci or replisomes, while the binding of fluorescent proteins to DNA, used to visualize genetic loci, may perturb replication or segregation of a locus (Possoz et al., 2006; Woldringh and Nanninga, 2006).
It is possible to zoom into a particular locus of the map to visualize dense areas in more details.
To visualize integration loci and their homologous regions microscopically, we used BACs covering the mapped chromosomal loci as FISH probes.
In summary, we developed novel tools to visualize chromosome loci in live cells with variant signal-to-noise ratios and signal intensities, and therefore broaden the adaptability of CRISPR/Cas9 system based imaging methods.
One hundred ng/ml of AT was added when binding of TetR-CFP to tetO arrays was used to visualize genetic loci.
