Buffer Series——Improve the efficiency of nucleic acid detection

When a certain amount of acid and alkali is added to some solutions, or a small amount of water is added to dilute, the solution has the effect of hindering the pH change of the solution, which is called buffering, and such solutions are called buffering solutions. Adding a small amount of strong acid or alkali to the buffer solution will not change the pH value of the solution much, but if the amount of acid and alkali is large, the buffer solution will lose its buffering effect. This shows that its buffer capacity is limited.

Buffer solutions have many uses. Among them, it plays an important role in maintaining the normal pH value and normal physiological environment of organisms. Most cells can only move in a narrow pH range, and a buffer system is required to resist the occurrence of metabolic processes. pH changes. There are three main pH buffer systems in organisms. They are protein and bicarbonate buffer systems. The weight of each buffer system is different in various types of cells and organs. For example, human blood contains multiple buffer pairs such as dihydrogen phosphate-hydrogen phosphate, carbonic acid-sodium bicarbonate, etc., to maintain the blood pH between 7.35 and 7.45 to maintain enzyme activity.

In biochemical research work, buffer solutions are often used to maintain the pH of the experimental system. The change of pH value of the solution system of research work often directly affects the effectiveness of our work. If the pH value of the extraction enzyme experiment system changes or changes too much, the enzyme activity will decrease or even be completely inactivated. In biochemical experiments or research work, buffer reagents should be carefully selected, because sometimes the factor that affects the experimental results is not the pH value of the buffer, but a certain ion in the buffer.

Application of buffer reagents in nucleic acid detection: buffer reagents use the pH change before and after the amplification reaction to make the amplification results can be distinguished by the naked eye, avoiding the use of complicated instruments and improving the detection efficiency. Fluorescence technology is commonly used in many detection methods of nucleic acid sequences and nucleic acid biological signals. However, these technologies require complicated equipment, which is heavy and expensive. The buffer solution can provide nucleic acid amplification colorimetric reaction. The pH-sensitive dye-based colorimetric method completely gets rid of the dependence on large-scale instruments, so that the results of the amplification reaction can be visualized with the naked eye, which is more suitable for on-site rapid detection needs. The principle is to use the production of pyrophosphate during the nucleic acid amplification reaction to cause the initial pH of the reaction solution to change to acidity, thereby causing the color of the pH sensitive dye in the reaction system to change. The buffer can promote the amplification reaction during application, can realize the visual detection of nucleic acid amplification in a shorter time and more efficiently, and can be used to realize fluorescence detection and colorimetric detection. At the same time, in nucleic acid detection, protein extraction needs to be stored in a buffer, and DNA is a relatively stable molecule. However, the introduction of nucleases into the DNA solution should be avoided because such enzymes can cause DNA degradation. Genomic DNA is composed of super large DNA molecules, so it is relatively fragile and easily damaged. DNA is easily hydrolyzed by acid when stored in water, so it should be stored in a buffer.

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