Hoechst dye

Incorporation of BUdR for replication banding permits direct detection of bands and signal when stained with the Hoechst dye. 

Ultraviolet Hoechst dyes, DAPI, Indo-1, AMCA,... 

Several types of fluorescent stain are available for the analysis of DNA their characteristics make them suitable for different applications (Table 8.1). The most specific stains (e.g., DAPI and the Hoechst dyes, which stain specifically for AT groups on DNA) require the use of a laser with significant ultraviolet (UY) output. Hoechst dyes as well as a newly developed far-red dye called DRAQ5 (alone of all the current DNA-specific stains) also penetrate the outer... 

Use a cell-permeant Hoechst dye, such as Hoechst 33342 (Molecular Probes, Eugene, OR). Permeablization of GFP-expressing cells with detergents may alter GFP localization and reduce fluorescent signal. 

Stain slides with a solution of Hoechst dye for 5 min, remove excess and carefully place coverslip onto slide. Seal edges with clear nail varnish (or equivalent) to minimize evaporation of water. 

To allow visualization of the location of the internalized complexes in relation to the cell nuclei, add 1 pi per well of the cell-permeable nuclear counterstain Hoechst 33342 (1 mg mh stock solution this results in a final Hoechst concentration of 5-lOpg/ml). Incubate for 15-20 min and observe with 350/461 nm blue fluorescence filters for Hoechst dyes (results are shown in Figs. 6a and b). 

Do not use phosphate-containing buffers, since Hoechst dyes precipitate in the presence of phosphates. 

Cyanine dyes and phenanthridine dyes, ethidium bromide and propidium iodide DAPl, Hoechst dyes , acridine orange, 7-AAD and hydroxystilbami-dine. 

SYBR dyes for sensitive detection in gels and blots, Chemically reactive SYBR dyes for bioconjugates. The three classes of classic nucleic acid stains are Intercalating dyes (ethidium bromide and propidium iodide), Minor-groove binders, (DAPI and the Hoechst dyes ). Miscellaneous nucleic acid stains with special properties (acridine orange, 7-AAD and hydroxystilbamidine). 39... 

There is evidence that poly(ADP-ribosyl)ation, in response to DNA damage (review [1]), permits changes in chromatin structure [2] which may facilitate the activity of repair enzymes [3] and become a major factor in the control of cell-cycle traverse [4], This paper explores the role of chromatin structure and poly(ADP-ribosyl)ation (using the inhibitor, 3-aminobenzamide 3AB [5]) in the responses of human cells to either a DNA specific ligand (Hoechst dye 33341 Ho33342) or X-radiation. Cells derived from an ataxia telangiectasia (A-T) patient have been included in the study to provide a h) ersensitive control (reviews [6, 7]) in which anomalous cell survival and cell-cycle responses [8-10] to radiation may reflect a primary defect in chromatin structure [11,12]. 

The detergent tends to increase backgrcnmd fluorescence of the Hoechst dye. 

It is particularly important to soak for extended periods because the significant thickness of the agarose means that removal of reagents is diffusion limited The detergent tends to increase background fluorescence of the Hoechst dye. 

For the total DNA content, the digested constructs were mixed with the Hoechst dye solution. The fluorescence emissions were read by using an absorbance reader. 

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