Principle of RNA extraction and tissue fragmentation

In the field of molecular biology research, RNA plays a crucial role as a transmitter of genetic information. However, it is not easy to extract RNA efficiently and accurately from complex biological tissues. Among them, tissue fragmentation is a key step in the RNA extraction process, and its principles and methods are directly related to the extraction efficiency and purity of RNA.

Principle of RNA extraction tissue fragmentation

1. Main PrincipleIn the process of RNA extraction, tissue fragmentation is one of the key steps. The principle is to quickly lyse cells by using ingredients in specific reagents (such as guanidine isothiocyanate, phenol, etc.). These components have a powerful protein denaturation effect, which can dissolve proteins and cause the secondary structure of the protein to disappear, thereby promoting the dissociation of the nucleoprotein soma. This process allows RNA that was originally closely bound to the protein to be released and enters the solution state.

2. Specific components and functions1. Guanidine isothiocyanate: This is a powerful protein denaturant that can quickly break cells and inhibit the nuclease activity released by cells while maintaining RNA integrity. It mainly achieves the purpose of lyzing cells by dissolving proteins and causing the secondary structure of proteins to disappear. 2. Phenol: Phenol can effectively denature proteins, and when used in combination with chloroform, it can enhance the inhibition of endogenous and exogenous RNase. This helps protect the RNA from degradation and ensures that the extracted RNA has a high purity.

3. Auxiliary steps and ingredientsIn the RNA extraction process, in addition to using protein denaturant for tissue fragmentation, a series of auxiliary steps and ingredients also need to further purify and extract RNA. For example: 1. Chloroform extraction: Chloroform, as an organic solvent, can quickly destroy cells and remove fat-soluble impurities in the homogenate. At the same time, it can also inhibit the activity of RNA enzymes and denature the protein, making it easy to remove by centrifugation. This step helps separate RNA from impurities such as DNA and protein. 2. Isopropanol precipitation: Isopropanol is used to increase the local density of RNA and make it precipitate out of solution. The precipitated RNA can be collected by centrifugation to obtain a relatively pure RNA product.

4. PrecautionsWhen RNA extraction tissue is broken, the following points should be paid attention to: 1. During the operation, try to create an environment without RNA enzymes, including removing contamination of exogenous RNA enzymes and inhibiting endogenous RNA enzyme activities. This helps reduce the risk of RNA degradation. 2. Sample treatment should be prompt and fast to avoid long-term exposure to room temperature, causing RNA degradation. 3. The reagents and vessels used should be sterile and free of RNA enzyme contamination.

The principle of RNA extraction and tissue fragmentation mainly relies on high concentrations of protein denaturants (such as guanidine isothiocyanate and phenol in Trizol reagents) to quickly cleave cells and release RNA. High purity RNA products can be further purified and extracted by a series of auxiliary steps and the treatment of ingredients.

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