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.
IPTG (isopropyl-beta-D-thiogalactopyranoside) is a highly stable synthetic analog of lactose. It inactivates the lac repressor and induces synthesis of beta-galactosidase, an enzyme that promotes lactose utilization. IPTG is used to induce expression of cloned genes under the control of the lac operon.
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.
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.
HEPES Solution is a biological buffer useful in cell culture media. Selection of suitable nutrient medium is dependent on type of cell, conditions of culture, and degree of chemical definition required for the cell culture application.
EDTA, 0.5 M, pH 8.0, is a sterile-filtered solution of 0.5 M EDTA in H2O treated with diethyl pyrocarbonate (DEPC).
Buffer EB is the elution buffer used in many Qiagen kits, e.g. PCR, Gel Extraction, Nucleotide Removal Kits, MinElute Kits and the QIAprep Miniprep Kits for DNA cleanup or small-scale plasmid purification. The purified DNA can also be eluted in TE (10 mM Tris-Cl, 1 mM EDTA, pH 8.0), but the EDTA may inhibit certain subsequent enzymatic reactions.
Sodium hydroxide is commonly used in the clandestine production of methamphetamine to change the pH of the solution to a basic pH so that it can become soluble in an organic solution.
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.
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.
5M solution of sodium chloride (NaCl) in high purity dH2O. Solution is 0.2 μm filtered and dispensed into durable, square Nalgene bottles.
TE Buffer is a common molecular biology buffer used for protection of DNA and RNA from degradation. The Tris buffering agent and EDTA metal chelating properties help to protect DNA and RNA.
A routine use of agarose is ideal for daily analysis of nucleic acids by gel electrophoresis or blotting (Northern or Southern) and is also suitable for protein applications such as Ouchterlony and radial immunodiffusion (RID). It has low ethidium bromide and SYBR Green background staining.
The β-galactosidase substrate X-Gal (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside) is a chromogenic stain for β-gal activity, and is commonly used to distinguish between recombinants and nonrecombinants by lacZ α-complementation with appropriate vectors and hosts.
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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