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.
MCLAB's high-purity 30% acrylamide/bis-acrylamide solution is a safer alternative to handling powdered acrylamide.
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.
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.
MCLAB’s 7M Guanidine-HCl Solution is a ready-to-use solution of guanidine hydrochloride, which can be easily diluted and pH-adjusted to any concentration below 7M.
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.
Phosphate-buffered saline (PBS) is a balanced salt solution that is used for a variety of cell culture applications, such as washing cells before dissociation, transporting cells or tissue samples, diluting cells for counting, and preparing reagents. PBS is formulated without calcium and magnesium for rinsing chelators from the culture before cell dissociation.
TAE buffer is the most commonly used buffer for DNA agarose gel electrophoresis but is also used for non-denaturing RNA agarose gel electrophoresis1. Double-stranded DNA tends to run faster in TAE than in other buffers but can also become exhausted during extended electrophoresis.
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.
Linear acrylamide is widely used as one of the coprecipitants which are inert substances used to aid recovery of nucleic acids before alcohol precipitations.
25% Triton X-100 is a commonly used detergent in laboratories.
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.
TWEEN 20 helps to prevent non-specific antibody binding in enzyme immunoassay as a washing agent (e.g. Western blots and ELISAs). In this major application, it is dissolved in Tris-Buffered Saline or Phosphate buffered saline at dilutions of 0.05% to 0.5% v/v.
Used to select for ampicillin resistance in mutated and transformed cells.
TRITON X-114 is used in biochemical applications to solubilize and separate proteins due to its low cloud point (23 °C).
DEPC-treated Water is deionized, high quality, molecular biology grade water.
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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