Agarose Gel Electrophoresis. Pouring of Agarose Gel, Preparation of samples, Gel electrophoresis, Ethidium bromide staining. - [Read Agarose Gel Electrophoresis PDF]
The most convenient and commonly used method to visualize DNA in agarose gels is staining with the fluorescent dye ethidium bromide. Ethidium bromide can be used to detect both singleand double-stranded nucleic acids (both DNA and RNA). However, the affinity of the dye for single-stranded nucleic acid is relatively low and the fluorescent yield is comparatively poor. - [Read Detection of DNA in Agarose Gels Protocol]
This simple cell culture-based cytotoxicity test (in which cell viability is determined by uptake of the dyes ethidium bromide and fluorescein acetate) has been developed as a general test for acute toxicity. - [Read LS-L929 Cytotoxcitiy Test]
Procedure permits the isolation of at least 5 µg of total RNA from a sample of purified mouse splenic B lymphocytes. The quality of the RNA is assessed by separation of an aliquot through 1% agarose and staining with ethidium bromide as described in AfCS protocol Visualization of RNA
Preparations on 1% Agarose Gels. The isolated RNA is used for analysis of gene expression by microarray technology.
analysis of gene expression by microarray technology. - [Read Preparation of B-Lymphocyte RNA for Microarray Analysis Protocol]
This protocol describes the preparation of concatamerized oligonucleotides and their coupling to cyanogen-bromide-activated Sepharose. The procedure uses a commercially activated resin, which can be purchased as a lyophilized powder. Keith Brocklehurst et al. - [Read Preparation of DNA Affinity Resin - Subscription Required]
A solution containing plasmid DNA, saturating amounts of ethidium bromide, and CsCl (44% w/v) is layered between two solutions of lesser (35% w/v CsCl) and greater density (59% w/v CsCl). During centrifugation to equilibrium, the closed circular plasmid DNA and linear DNAs form bands at different densities. - [Read Purification of Closed Circular DNA by Equilibrium Centrifugation in CsCl-Ethidium Bromide Gradient]
Solutions containing plasmid DNA are adjusted to a density of 1.55 g/ml with solid CsCl. The intercalating dye, ethidium bromide, which binds differentially to closed circular and linear DNAs, is then added to a concentration of 200 mu;g/ml. During centrifugation to equilibrium, the closed circular DNA and linear DNAs form bands at different densities. - [Read Purification of Closed Circular DNA by Equilibrium Centrifugation in CsCl-Ethidium Bromide Gradients]
Separation of RNAs according to size is the first stage in northern blotting and hybridization. The method described in this protocol uses formaldehyde to denature the RNA, ethidium bromide to stain it, and electrophoresis through agarose gels containing 2.2 M formamide to separate the resulting formaldehyde-RNA-ethidium adducts. - [Read Separation of RNA According to Size: Electrophoresis of RNA through Agarose Gels Containing Formalde]
Method is used to assess (roughly) the integrity of total RNA samples by visualization of discreet 18S and 28S ribosomal RNAs. Total RNA is separated by electrophoresis through a 1% agarose gel containing 1.3 ìM ethidium bromide. Binding of the ethidium bromide to the RNA allows visualization of the separated RNA molecules when the gel is exposed to ultraviolet (UV) light. - [Read Visualization of RNA Preparations on 1% Agarose Gels Protocol]