A ribonucleic acid protecting agent, kit, application and preservation method

Abstract

The invention provides an RNA protective agent, a kit containing the protective agent, application and a preservation method of RNA. The protective agent contains a trehalose solution with a concentration of 0.27M-1M. The protective agent provided by the invention can achieve the effects of long preservation time, temperature insensitivity and wide range of use temperature, an extracellular RNA sample treated by the protective agent has no influence to cDNA synthesis, PCR reaction and enzyme activity in sequencing and bank building. The protective agent provided by the invention can efficiently solve the RNA degradation, pollution and other problems encountered during long-term preservation or use of exRNA, and especially has important significance for preservation, repeated use and long-distance transportation of extracellular microRNA.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to an RNA protection agent, a kit, an application and a preservation method. Background technique [0002] Ribonucleic acid (abbreviated as RNA, that is, Ribonucleic Acid), the genetic information carrier that exists in biological cells and some viruses and viroids, RNA is a long chain molecule formed by ribonucleotides condensed through phospholipid bonds, and can participate in The various life activities of cells are the key research objects in the fields of medicine, biology and pharmacy. [0003] With the in-depth study of RNA, it is found that cells with vital activities can secrete specific RNA from the cell to the circulatory system or specific tissue microenvironment, and this type of RNA is defined as extracellular RNA (exRNA). Due to the obvious difference between exRNA and common RNA in cells, exRNA can play various potential biological roles as a signal. At present, t...

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Problems solved by technology

However, exRNA is often degraded by RNase after long-term storage, freeze-thawing and purification, which brings great difficulties to the widespread use, storage and transportation of exRNA

[0004] In the practical application of RNA, the preservation of RNA depends on pure water without RNase or an aqueous solution with chelating agents such as EDTA added to dissolve the RNA and store it in an ultra-low temperature freezer at -80°C for one year. For example, irreversible RNA fragmentation will be caused during freezing at room temperature to -80°C, and physical damage will destroy the original biological activity of microRNAs when returning to room temperature at -80°C.

[0005] There are currently several commercial reagents on the market for preserving biological samples containing RNA, such as RNA later from LifeTech, RNA stable from Biomatrica, etc., whether they are stored in liquid or solid state, the enzyme activity reaction inhibitors and denaturation in these reagents Reagents can protect RNA samples from RNase degradation, but the disadvantage of these reagents is that the original RNA samples need to be purified before being used in subsequent biological reactions, and the purification process will further lead to the loss of exRNA samples, and the purified exRNA is especially The solid-state storage of microRNAs is of great significance for the long-distance transportation and repeated use of microRNAs

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