The loading dye increases the density of the sample and adds colours to the sample, thereby simplifying the loading process. The solution contains dyes that, in a electric field, move toward the anode at predictable rates.
Description
The loading dye increases the density of the sample and adds colours to the sample, thereby simplifying the loading process. The solution contains dyes that, in a electric field, move toward the anode at predictable rates.
In 1% agarose gels, bromophenol blue migrates with a 300 bp linear double-stranded DNA fragment, whereas xylene cyanol FF migrates at approximately the same rate as linear double-stranded DNA 4 kb length. These relationships are not significantly affected by the concentration (0.5 to 1.4%) of agarose in the gel.
Loading: The loading dye suits well for DNA samples dissolved either in water or in EDTA-containing buffer (as TE buffer). If DNA markers are not prediluted with the Loading dye solution, then mix. The loading buffer is 6x concentrated, that means you have to use it 5:1.
For DNA markers, apply 0.1 µg per 1 mm of agarose gel lane width. Often 1µg of marker is used in one electrophoresis run but it depends on the size of your gel and the comb.
Applications
Preparation of DNA ladders, markers, and samples for loading on agarose or polyacrylamide gels.
Recommended Storage Conditions: Store at room temperature or at 4 °C up to 12 months. For longer periods, store at -20 °C.
Low concentration solutions of DNA are prone to DNA degradation or other loss. It is critical that optimal methods are employed for DNA suspension and long-term storage.
Bovine Serum Albumin (BSA) has many uses as a carrier protein and as a stabilizing agent in enzymatic reactions. MCLAB’s Ultrapure BSA is a "non-acetylated" BSA, pure enough to use when the integrity of DNA or RNA is essential. It has been tested for DNase, RNase, endonuclease, protease, peroxidase, and alkaline phosphatase activity, and assayed for fluorescence background.
CHAPSO (3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate) is a zwitterionic detergent used to solubilize proteins. Due to its low light absorbance in the ultraviolet region of the electromagnetic spectrum, CHAPSO is useful for researchers monitoring ongoing chemical reactions or protein-protein binding with UV/Vis spectroscopy.
Brij-35 (30% Solution) is a nonionic polyoxyethylene surfactant that is most frequently used as a component of cell lysis buffers or as a surfactant in various HPLC applications.
Ammonium Persulfate (APS) is an oxidizing agent that is used with TEMED to catalyze the polymerization of acrylamide and bisacrylamide to prepare polyacrylamide gels for electrophoresis.
Ammonium acetate is a widely used reagent in molecular biology and chromatography. Suitable applications include the purification and precipitation of DNA and protein crystallization. Ammonium acetate is commonly used in HPLC and MS analysis of various compounds, such as oligosaccharides, proteins, and peptides.
Acetylated BSA is used as a carrier protein and an enzyme stabilizer in which the absence of contaminants such as nucleases and proteases is vital. The acetylation process inactivates any trace of active nuclease activity. While the acetylation process does change BSA's binding characteristics and makes it inhibitory in PCR reactions, it is normally added to restriction digests.
EDTA, 0.5 M, pH 8.0, is a sterile-filtered solution of 0.5 M EDTA in H2O treated with diethyl pyrocarbonate (DEPC).
Linear acrylamide is widely used as one of the coprecipitants which are inert substances used to aid recovery of nucleic acids before alcohol precipitations.
Tris Buffered Saline (TBS) is an isotonic, non-toxic buffer used in some biomedical techniques to maintain pH within a relatively narrow range. TBS can shift pH relatively drastically as temperature changes, with the pKa declining approximately 0.03 units per degree Celsius rise in temperature.
TWEEN-40 is a class of emulsifiers used in some pharmaceutical and food preparations. It is often used in cosmetics to solubilize essential oils into water-based products. Polysorbates are oily liquid derived from PEGylated sorbitan (a derivative of sorbitol) esterified with fatty acids.
MCLAB's RNA Storage Buffer, with optimized reagents and pH value, provides better preservation and greater RNA stability for valuable RNA samples than the standard EDTA and TE Buffer or just water.
25% Triton X-100 is a commonly used detergent in laboratories.
D-(+)-Glucose is a common natural sugar involved in processes such as energy production, glycosylation, and formation of glycans that provide structure to cells. Glucose is involved in a detrimental process in cells called glycation. Glucose is used as a supplement for cell culture and in numerous cellular processes and molecular biology applications.
5M solution of sodium chloride (NaCl) in high purity dH2O. Solution is 0.2 μm filtered and dispensed into durable, square Nalgene bottles.
SDS, sodium dodecyl sulfate is an anionic surfactant. SDS is commonly used in laboratory as a component of buffer for cell lysis, cell lysis during DNA extraction, and mostly in SDS-PAGE running buffer. Indeed, SDS is an anionic detergent applied to protein samples to linearize proteins and to impart a negative charge to linearized proteins.
The loading dye increases the density of the sample and adds colours to the sample, thereby simplifying the loading process. The solution contains dyes that, in a electric field, move toward the anode at predictable rates.
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