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.
Description
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. Dilution of the concentrated TAE buffer produces a 1× TAE buffer with 40 mM Tris-acetate and 1 mM EDTA, pH 8.3. The 1× TAE buffer is used both in the agarose gel and as a running buffer.
Application
Casting and running buffer for agarose gel electrophoresis of nucleic acids.
Notes
0.4M Tris base, 0.4M acetate, 10mM EDTA, pH 8.0.
Recommended Storage Condition: Room temperature
DEPC-treated Water is deionized, high quality, molecular biology grade water.
25% Triton X-100 is a commonly used detergent in laboratories.
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.
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.
CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate ) is a zwitterionic detergent used to solubilize proteins. It is used as a non-denaturing solvent in the process of protein purification and is especially useful in purifying membrane proteins.
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.
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.
EDTA, 0.5 M, pH 8.0, is a sterile-filtered solution of 0.5 M EDTA in H2O treated with diethyl pyrocarbonate (DEPC).
Reported use in all phases of detergent compounding and aqueous processing in the textile and paper industries, in industrial metal cleaners, acid cleaners, floor cleaners, detergent sanitizers and waterless hand cleaners.
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.
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.
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.
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 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.
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.
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.
Čeština
Polski
