mRNA quantitative detection method

Abstract

The invention discloses an mRNA quantitative detection method. The method comprises the following steps: preparing standard mRNA color plates through optical film biological sensor chip preparation, probe synthesis and lattice, standard mRNA color plate sample preparation, cDNA synthesis and DIG label, hybridization, precipitation and coloration; preparing the sample to be detected through the optical film biological sensor chip preparation, probe synthesis and lattice, total RNA extraction of samples, cDNA synthesis and DIG label, hybridization, precipitation and coloration; and comparing the color by utilizing the colors presented by the mRNA of the sample and standard mRNA to determine the content of mRNA in the samples to be detected. In the method of the invention, the detection precision is high, which is improved by 100 thousand times compared to that of the agarose denaturing gel detection method; the detection speed is high, and the results can be detected in a short time of only 30 minutes, which takes less than 10 hours than the Northern hybridization method; the detection results can be directly read by naked eyes with no need of using equipment; the application prospect is wide, and can be generally applicable to biological mRNA quantization detection.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to a method for directly quantifying mRNA content in a sample by using an optical film biosensor chip, that is, an mRNA quantitative detection method. Background technique [0002] Quantitative detection of mRNA in samples plays an important role in molecular biology techniques. For example, the synthesis of cDNA probes in microarray analysis, the determination of RNA concentration before Northern hybridization (hybridization of RNA and DNA), the construction of cDNA library, RT-PCR (reverse transcriptase PCR) reaction and PCR differential display experiments are all involved. Quantitative detection of mRNA. Traditional mRNA quantitative detection methods include spectrophotometric detection methods, agarose denaturing gel detection methods, Northern hybridization detection methods, etc. [0003] The spectrophotometric detection method is the most c...

AI Technical Summary

Problems solved by technology

Although this method is easy to operate, the disadvantage is that during the RNA preparation process, free nucleotides, proteins, and common pollutants will generate a large number of interference signals, and it is difficult to separate the DNA, rRNA, and tRNA in the sample from the mRNA by absorbance. Accurately distinguish them, resulting in large errors in measurement results; and the relative sensitivity and accuracy of the detection results of this method are not high

However, the disadvantages of this method are: there are more requirements on the sample volume, otherwise the accuracy will be affected; when the total RNA amount in the sample is less than 1 mg, the detection result cannot be obtained

But the disadvantages of this method are: it still lacks precision, it takes a long time, and it also has the risk of radioactive contamination

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