RNA colorimetric quantitative detection kit product manual

The main purpose

The YIJI RNA colorimetric quantitative detection reagent is a technical method for determining the absolute content or concentration of RNA in a sample by a specific binding reaction of bromophenylhydrazine with RNA by means of a change in the absorption peak of visible light wavelength. It is suitable for cellular or tissue RNA analysis of various prokaryotic and eukaryotic organisms as well as RNA content analysis in the environment. The product is ready to use, stable performance, simple and convenient, no sample purification, accurate quantitative.

technical background

P-bromophenylhydrazine (PBPH) has ribosephosphate depurinated with RNA, and defurther dephosphoric acid and dehydrated furfural, which is extracted by acidification solvent to carry out specificity. The reaction forms a stable, reproducible yellow complex. At the same time, it effectively limits the interference caused by the deoxyribose released by DNA and the reaction of monosaccharide molecules such as glycogen, methyl glucoside, levulose and dextrose in the sample. The p-bromoquinone method is more specific than the orcinol reaction. Quantitative analysis of RNA by visible light (450 nm) absorption.

product content

YIJI hydrolyzate (Reagent A) ml

YIJI acidification solution (Reagent B) ml

YIJI Extract (Reagent C) ml

YIJI reaction solution (Reagent D) ml

YIJI Stabilizer (Reagent E) ML

YIJI standard solution (Reagent F) ml

Product manual 1 copy

storage method

Preserve YIJI standard solution (Reagent F) in -20 °C refrigerator, the rest is stored in 4 ° C refrigerator; reagent is highly corrosive, pay attention to safe operation, and avoid light, effectively guarantee June

User-supplied

15 ml conical centrifuge tube: container for sample handling

Dry thermostat or electric furnace: for reactant incubation

Benchtop centrifuge: for sample handling

3 ml glass cuvette: 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 450nm, and set zero
• Prepare 6 1.5 ml tubes, labeled 1 to 6
• Prepare standard solution according to the following table
• Put it in the ice tank for use, avoiding light

Second, colorimetric determination

• Prepare at least 7 new 15 ml conical centrifuge tubes as assay tubes: label 6 standard tubes and 1 sample tube to be tested
• Add xx μl of the above-prepared YIJI standard solution (Reagent F) to the corresponding standard measuring tube (note: concentration and tube number are in place)
• Add 500 μl of the sample to be tested into the sample tube
• Add xx microliters of YIJI hydrolyzate (Reagent A) and mix separately.
• Boil for 30 minutes or put in a 100 °C dry thermostat for 60 minutes (note: carefully open the lid during heating)
• Allow to stand at room temperature until the tube temperature drops to room temperature (15 to 30 minutes)
• Add xx ml of YIJI acid solution (Reagent B) to all measuring tubes.
• Add xx ml of YIJI extract (Reagent C)
• Boil for 60 minutes or put in a 100 °C dry thermostat for 3 hours (note: carefully open the lid during heating)
• Allow to stand at room temperature until the tube temperature drops to room temperature (15 to 30 minutes)
• Add xx ml of YIJI extract (Reagent C)
• Vortex oscillation for 15 seconds
• Place in a benchtop centrifuge for 15 minutes at a speed of 10,000g
• Carefully remove 1 ml of the upper liquid phase into a new 15 ml conical tube
• Add xx ml of YIJI reaction solution (Reagent D) and mix
• Incubate for 1 hour at room temperature to avoid light
• Add xx ml of YIJI Stabilizer (Reagent E) and mix
• Transfer to a 3 ml glass cuvette
• Put in the spectrophotometer reading: get absorbance reading
• Construct standard curve: ordinate (Y axis) is the light absorption unit (OD); abscissa (X axis) is the standard RNA content (microgram)
• Obtain the corresponding RNA concentration (microgram) of the sample according to the standard curve
• Sample actual RNA concentration:

Precautions

• This product is 10 sample operations
• This product is tested from 9 to 150 micrograms.
• Wear gloves when handling
• Standard sample measurement is only required once during system operation
• Reagents are corrosive and strictly observe operational safety
• The sample to be tested does not need to be purified, including deproteination, de-sugar removal, DNA 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 biological macromolecular quantitative analysis reagent products.

Quality Standard

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

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