Kit for ribonuclease protecting experiment

Abstract

The invention relates to a ribonuclease protecting experiment kit. It includes kit body and many reagent containers. The many reagent containers are fixed in the kit body. The kit body includes hybridization, RNA enzyme digestion, probe dilution, and sample adding buffer solution containers. The hybridization bugger solution is the following mixture: 40mmol / L PIPES(pH6.8), 1mmol / L EDTA(pH8.0), 0.4mol / L NaCl, 1mmol / L CTAB, and deionization formamide with 80% volume ratio. The RNA enzyme digestion bugger solution is the following mixture: 0.3mol / L NaCl, 5mmol / LEDTA, 10mmol / L Tris-HCl(pH7.4) and 50mg / L glycogen. The invention has the advantages of high sensitivity, good repeatability, and low production cost. And it can be applied to non radioactivity biotin sign probe.

Description

technical field [0001] The invention relates to reagents used in gene detection, in particular to kits used in messenger ribonucleic acid (mRNA) analysis. Background technique [0002] The expression of a gene needs to go through transcription (mRNA production), translation (protein synthesis), modification and other processes, and finally form a protein molecule with biological activity. mRNA detection is the most important and commonly used method to study gene expression and function. Currently commonly used mRNA detection methods include Northern blot, RT-PCR and ribonuclease protection assay (Ribonuclease protection assay, RPA). [0003] The Northern blot method is one of the classic methods for mRNA detection, which can semi-quantitatively analyze the level of mRNA, but the method is complicated to operate, takes a long time, requires high integrity of the specimen, requires a large amount of samples, and cannot be detected at the same time Multiple target genes; Qua...

AI Technical Summary

Problems solved by technology

However, using existing kits requires a long hybridization time, and during the experiment of digesting unhybridized single-stranded RNA, the precipitation of undigested protection fragments is not sufficient, which increases the difficulty of observation

[0005] In addition, there are very few reports on the use of this method in China. One of the important reasons is that there is currently no convenient and economical kit available in China, and imported reagents are expensive.

On the other hand, most of the existing kits are only suitable for the use of radioactive isotope-labeled probes, which have a certain impact on the environment and the health of researchers, and have high requirements for the experimental site and its protection, thus limiting Promotion of the technology in the country

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