Why Isopropanol is Used to Extract RNA

Why is isopropanol used to extract RNA?

Isopropanol (Isopropanol) is a common organic solvent, which is widely used in biological experiments, especially in the process of RNA extraction. Many people may wonder why isopropanol is used to extract RNA and not just other solvents. This article will explore the role and importance of isopropanol in RNA extraction in detail.

RELATIONSHIP OF ISOPROPANOL CHEMICAL PROPERTIES WITH RNA EXTRACTION

Isopropanol is a less polar solvent whose chemical properties make it effective in helping the RNA to precipitate out of the cell solution. In the process of RNA extraction, the main role of isopropanol is to reduce the polarity of the solution, so that the RNA molecules are no longer dissolved with the aqueous phase, thereby forming a precipitate. The solubility of RNA molecules in aqueous solution is high, while the solubility in isopropanol is significantly reduced, which allows RNA to be isolated from the sample.

Mechanisms that promote RNA precipitation

When extracting RNA, the use of isopropanol can reduce the ionic strength of the solution to some extent. When isopropanol is added to the cell lysate, the hydrated ions are removed and the original hydration on the surface of RNA molecules is weakened, resulting in the aggregation of RNA molecules and the formation of precipitation. This precipitation process is critical for RNA purification because it effectively removes proteins, lipids, and other impurities dissolved in the aqueous phase, resulting in relatively pure RNA.

Isopropanol Volatility and RNA Protection

Isopropanol is less volatile and evaporates more quickly after use, avoiding the problem of RNA degradation with prolonged exposure to water. Isopropanol solution can reduce the activity of RNA degrading enzymes (such as RNase) and protect RNA from the external environment. This is essential to maintain the integrity of the extracted RNA, especially for downstream applications, such as qPCR, Northern blot and other analysis.

Isopropanol in RNA Extraction Advantages

The use of isopropanol can improve the efficiency of RNA extraction compared to other solvents, such as ethanol. Under the same conditions, isopropanol was able to precipitate RNA more rapidly, reducing the operating time and increasing the yield. Moreover, the application of isopropanol in RNA extraction does not lead to excessive dissolution of RNA, so its precipitation effect is more ideal.

Summary

As a key reagent in RNA extraction, isopropanol has excellent solvent properties, which can effectively promote the precipitation of RNA, improve the extraction efficiency, and protect RNA from degradation. In the process of RNA extraction, the role of isopropanol is indispensable, which provides important technical support for biological research and molecular biology experiments. Therefore, the answer to the question of why isopropanol is used to extract RNA lies in its unique chemical properties and its protective effect on RNA, making it an indispensable solvent in the RNA extraction process.