What is nucleic acid isolation?
What is nucleic acid isolation?
The isolation of nucleic acids mainly refers to the separation of nucleic acids from biological macromolecules such as proteins, polysaccharides, and fats. The following principles should be followed when isolating nucleic acids: ensure the integrity of the primary structure of nucleic acid molecules; exclude other molecular contamination. Steps of nucleic acid isolation and purification Most nucleic acid isolation and purification methods generally include several main steps, such as cell lysis, enzymatic treatment, separation of nucleic acid and other biological macromolecules, and nucleic acid purification.
Nucleic acid research is an important subject in modern biology and medical research. As the carrier of genetic information, nucleic acid is the material basis of gene expression. In addition to playing a very important role in the normal growth, development, reproduction and other life activities of organisms, it is closely related to abnormal conditions of life, such as tumorigenesis, radiation, etc. Injuries, genetic diseases, etc. are also closely related. Therefore, nucleic acid is an important subject in modern biology and medical research. Whether conducting research on nucleic acid structure and function, or conducting research on genetic engineering, protein engineering, etc., it is first necessary to separate and purify nucleic acid.
The traditional nucleic acid separation technology includes precipitation, centrifugation and other processes. These purification methods have complicated steps, time-consuming, low yield, contact with toxic reagents, and it is difficult to realize automatic operation; and the magnetic carrier microsphere separation technology can be well overcome. With these shortcomings, the rapid and efficient preparation of samples is an important direction for the development of nucleic acid purification methods in the future.
Introduction to nucleic acid extraction by magnetic bead method
Nucleic acid extraction by magnetic bead method is to lyse cells by cell lysate, and the nucleic acid molecules freed from the cells are specifically adsorbed to the surface of magnetic particles, while impurities such as proteins are not adsorbed and remain in the solution. After a certain time of reaction, under the action of a magnetic field, the magnetic particles are separated from the liquid, and the particles (ie, the magnetic bead-DNA mixture) are recovered, and then eluted with the eluent to obtain pure DNA.
The magnetic bead method does not require centrifugation, does not need to add a variety of reagents, is simple to operate, and meets the requirements of automated nucleic acid extraction. It is an important direction for the development of nucleic acid purification methods in the future.
Simple process of technical route
Sample lysis - specific binding of magnetic beads to DNA to be separated - washing of magnetic bead-DNA mixture - elution separation of magnetic beads and DNA to be separated (ie three steps of binding-washing-elution).
After the cells are lysed, magnetic beads are added to the lysing solution. When the pH value of the solution is less than 6.5, the magnetic beads selectively bind to the optimized DNA. At this time, the magnetic beads adsorbed with DNA are placed in a magnetic field and removed by the buffer. There are no adsorbed impurities (protein, etc.), and then the magnetic beads are placed in a buffer with a pH value of 8.5, and the purified DNA can enter the buffer.
The advantage of micro-nano magnetic bead nucleic acid purification
With the micro-nano magnetic bead nucleic acid purification reagent, the sample recovery and purification effect is optimized, and it can be directly used in subsequent experiments.
1. The automatic operation ensures the stability of the experimental results and avoids the differences and errors caused by manual operation.
2. The intelligent operating system strictly controls the contamination between the holes, and the disposable plastic extraction sleeve is used to avoid the contamination between different batches.
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