Diphenylamine DNA colorimetric quantitative assay kit product specification

The main purpose

The diphenylamine DNA colorimetric quantitative assay reagent is a technical method for determining the absolute content or concentration of DNA in a sample by means of a specific binding reaction of diphenylamine with DNA, by means of a change in the peak of the visible light wavelength. It is suitable for cell or tissue DNA of various prokaryotes and eukaryotes as well as DNA content analysis in the environment. The product is ready to use, stable performance, simple and convenient, no sample purification, accurate quantitative.

technical background

Diphenylamine has a function of specifically reacting with deoxyribose-transformed ω-hydroxylevulinyl aldehyde released after acid hydrolysis and desorption of DNA to form a blue complex. But does not react with the ribose of RNA. Quantitative analysis can be performed by absorption of visible light (600 nm). Thus, various limitations of the A260 assay are avoided, such as purification requirements for the sample, requirements for single-stranded duplexes of DNA, and UV UV requirements, as well as RNA interference.

product content

Buffer (Reagent A) 8 ml

Decomposition solution (Reagent B) 8 ml

Reaction solution (Reagent C) 25 ml

Stabilizer (Reagent D) 125 μl

Standard solution (Reagent E) 200 μl

Product manual 1 copy

storage method

The storage solution (Reagent D) and the standard solution (Reagent E) are stored in a refrigerator at 4 ° C, and the rest are stored at room temperature; the reaction solution (Reagent C) and the stabilizer (Reagent D) are corrosive, and the operation is safe. And avoid lighting, effectively guaranteeing June

User-supplied

1.5 ml centrifuge tube: container for sample handling

Constant temperature sink: for reactant incubation

Cuvette: a container for colorimetric analysis

Spectrophotometer: for sample colorimetric determination

Experimental procedure

• Preparation for measurement
• Prepare the sample to be tested and place it in the ice trough
• Set the spectrophotometer: wavelength 600nm, and set zero
• Prepare 9 1.5 ml centrifuge tubes labeled 1 to 9
• Prepare standard solution according to the following table
• Put it in the ice tank for use, avoiding light

Second, colorimetric determination

Before the start of the experiment, carefully remove 6 ml of the reaction solution (Reagent C) and 30 μl of the stabilizer (Reagent D) into a 15 ml conical centrifuge tube. After mixing, mark the reaction solution to avoid light. Then do the following.

• Prepare at least 10 new 1.5 ml centrifuge tubes as assay tubes: label 9 standard tubes and 1 sample tube to be tested
• Add 140 μl of buffer (Reagent A) to each standard and sample tube
• Add 10 μl of the above prepared standard solution (Reagent E) to the corresponding measuring tube (Note: Concentration and tube number are seated)
• Add 10 μl of the sample to be tested into the sample tube
• Add 150 μl of decomposition solution (Reagent B) to all assay tubes
• Incubate in a 70°C dry bath or a constant temperature bath for 20 minutes.
• Allow to stand at room temperature until the tube temperature drops to room temperature (15 to 30 minutes)
• Were added 600 l of a reaction solution containing (Reagent C) and the reaction fluid stabilizing solution (Reagent D) to the measuring tube in all
• Incubate for 5 hours in a 37 ° C incubator (dark room environment)
• Immediately move to a 1 ml cuvette and place in the spectrophotometer reading: Obtain absorbance readings
• Construct a standard curve: the ordinate (Y-axis) is the unit of absorption (OD); the abscissa (X-axis) is the standard DNA concentration (μg/10 μl)
• Obtain the corresponding DNA concentration of the sample according to the standard curve (μg/10 μl)
• Actual DNA concentration of the sample:

[According to the standard curve, obtain the corresponding DNA concentration (μg/10 μl) X sample dilution factor] ÷100=μg/ml

Precautions

• This product is 10 sample operations
• Wear gloves when handling
• Standard sample measurement is only required once during system operation
• Reagent C, Reagent D and reaction working solution to avoid light
• Be careful when handling, especially when using the reagent solution (Reagent C), stabilizer (Reagent D) and reaction working fluid .
• The sample to be tested does not need to be purified, including deproteination, RNA removal, etc.
• When calculating the actual concentration, don't forget the dilution factor of the sample to be tested.
• Wash the cuvette with absolute ethanol
• The company provides a series of DNA quantitative analysis reagent products

Quality Standard

• This product has been certified to be stable.
• This product has been identified and quantified accurately

F-Phenibut FAA

Carnitine, or trans. carnitine, is an amino acid, a quaternary ammonium cationic complex, which can be biosynthesized from both lysine and methionine and is involved in the metabolism of fat into energy in the body. Carnitine has two stereoisomerism: L-Carnitine, which is biologically active, and D-carnitine, which is non-biologically active. L-carnitine (L-carnitine) is an amino acid widely distributed in the liver, especially in myocardium and skeletal muscle. Most of the carnitine required by the body comes from meat and dairy products in the diet. [1]