The advantages and disadvantage of DNA extraction by spin column method
As one of the most basic experiments in molecular experiments, nucleic acid extraction is the first step in starting biological research. No matter the subsequent cloning, PCR, qPCR, library construction and sequencing, etc., the nucleic acid is required to proceed smoothly, and the quality of the extracted nucleic acid is also One of the key factors that determine the success or failure of downstream experiments.
Nucleic acid extraction can be divided into three types: solution extraction, column extraction, and magnetic bead purification. Solution extraction is a more classic extraction method, but it is more complicated and the quality of the extracted nucleic acid is also general. Column extraction is a relatively simple method for the isolation and purification of trace nucleic acids. The extraction quality is good, but it is not suitable for the extraction of large quantities of samples. Magnetic bead nucleic acid extraction has the advantages of simplicity, convenience, high throughput, and non-toxicity. Cell tissue samples are lysed by lysis solution, and the nucleic acid molecules freed from the samples are specifically adsorbed to the surface of magnetic particles, and impurities such as proteins are not adsorbed. remain in solution. Then let's introduce the best way to isolate and purify trace nucleic acids.
What is spin column extraction of nucleic acids?
Spin column extraction is a relatively simple method for the separation and purification of trace nucleic acids.
The released nucleic acid is specifically adsorbed on a specific silicon carrier. This carrier only has a strong affinity and adsorption force for nucleic acid, and basically does not adsorb other biochemical components such as proteins, polysaccharides, and lipids. Therefore, during centrifugation was thrown off the pillar. Then, the nucleic acid adsorbed on the specific carrier is eluted with the eluent, and the purified nucleic acid is obtained by separation.
Column extraction of nucleic acid has a lower cost, but better extraction quality, and is currently the mainstream extraction method in the market.
The working principle of DNA extraction by spin column method
The unique binding solution/proteinase K rapidly lyses cells and inactivates intracellular nucleases, and then genomic DNA is selectively adsorbed on the silica matrix membrane in the spin column in a high-salt state, and then through a series of rapid washing-centrifugation steps, inhibiting The cell metabolites, proteins and other impurities are removed by the compound removal solution and rinsing solution, and finally, the pure genomic DNA is eluted from the silica matrix membrane by the low-salt elution buffer.
1. Adopt a centrifugal adsorption column and buffer system. After the sample is lysed, DNA is bound to the silica membrane under high salt conditions and eluted from the silica membrane at low salt and high pH.
2. Destroy red blood cells. Usually, the difference in the membrane structure of red blood cells and white blood cells is used to first lyse red blood cells, and collect white blood cells after centrifugation.
3. Leukocyte fission denatures membrane proteins and nuclear proteins, and free DNA is usually denatured by ionic surfactants.
4. Removal of denatured proteins: Usually, protein precipitants are used to precipitate denatured proteins, leaving DNA in the supernatant.
5. Precipitate DNA from organic solvents (such as absolute ethanol) in a high-salt environment.
Advantages of DNA Extraction by Spin Column Method
The advantage of DNA extraction by spin column method is that it has higher purity than the traditional phenol chloride method, which is beneficial to RNA protection and can carry out micro-operation. With its low price and relatively convenient operation, it gradually replaces the traditional DNA extraction method. It is generally accepted by the market such as universities and research institutes.
Disadvantages of spin-column DNA extraction
The disadvantage of DNA extraction by spin column method is that it requires more samples and consumables, and is powerless for rare samples, especially in the fields of forensics and archaeology. At the same time, DNA extraction by spin column method requires repeated centrifugation, which is inconvenient for high-throughput and automated operations and is incompatible with the high-throughput and automation requirements of modern biological experiments, especially in the fields of gene diagnosis, disease detection, and transgenic detection. DNA extraction requires a large number of operators and equipment.
Professional manufacturer of Nucleic Acid (DNA & RNA) Extraction and Analysis products spin column supplier
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