In molecular biology experiments, DNA precipitation and extraction are fundamental steps in gene research, cloning technology, and diagnostic reagent development. During this process, the selection of biological buffering agents directly affects the purity, integrity, and experimental efficiency of DNA. Trimethylmethylaminoethanesulfonic acid (TES buffer), as an efficient buffering agent, has demonstrated significant advantages in the field of DNA manipulation due to its unique physicochemical properties and stability.
The chemical properties of TES: the scientific basis for buffering capacity
TES, The chemical name is 2- [(trihydroxymethyl) methylamino] -1-ethanesulfonic acid, and the sulfonic acid groups and multiple hydroxyl groups in its molecular structure endow it with excellent buffering ability. At room temperature, the pKa value of TES is 7.5, and the effective buffering range covers pH 6.8-8.2, which precisely matches the activity and DNA stability requirements of most biological enzymes. Compared to traditional buffer Tris (buffer range pH 7.0-9.0), TES exhibits stronger resistance to temperature changes and can effectively avoid the risk of low-temperature induced DNA degradation.
Triple core role in DNA extraction
1. Cell lysis and protein removal
During the cell lysis stage, TES inhibits the activity of DNA enzymes and protects genomic DNA from degradation by chelating divalent metal ions (such as Mg ² ⁺, Ca ² ⁺) in the solution. Its buffer system simultaneously maintains the optimal pH for proteases such as proteinase K (usually pH 7.5-8.0), accelerating the breakdown of cell membranes and protein degradation. Experiments have shown that the protein removal efficiency of the lysis system using TES buffer is improved by about 25% compared to traditional methods, laying a good foundation for subsequent purification steps.
2. Selective regulation of DNA precipitation
TES is a composite buffer solution composed of Tris hydrochloric acid, EDTA, and SDS (TES-EDTA-SDS), which achieves selective precipitation of DNA by adjusting the ionic strength of the solution. At high salt concentrations (such as 2.5M NaCl), the sulfonic acid groups of TES act through electrostatic shielding, promoting the repulsion of phosphate groups between DNA molecules and forming a soluble state; When the salt concentration drops below 0.5M, DNA aggregates and precipitates due to exposure of the phosphate backbone. This process has a repulsive effect on RNA and other impurities, resulting in DNA recovery purity of over 95%.
3. Enzyme activity protection
The protective effect of TES buffer on enzyme activity is particularly prominent in limiting enzyme cleavage reactions. Research has shown that the cutting efficiency of restriction enzymes (such as EcoRI) using TES reaction system is 18% -22% higher than that of HEPES buffer. Its stable pH environment reduces enzyme conformational changes, prolongs the linear period of enzyme cleavage reactions, and ensures the accuracy of gene editing experiments.
Conclusion
TES buffer exhibits irreplaceable advantages in DNA precipitation and extraction due to its excellent pH stability, metal ion chelation properties, and enzyme protection effect. By optimizing experimental conditions and exploring new application scenarios, TES will continue to drive molecular biology research towards more efficient and precise directions. For researchers, mastering the buffering mechanism and application techniques of TES is undoubtedly the key to improving the success rate of experiments.
TES buffer is not only used in cosmetics. Its buffering properties make it play an important role in protein analysis, enzyme activity determination, and cell culture. As an advantageous supplier of TES buffer, Desheng can provide high-purity and diverse types of buffer raw materials. Customers can complete one-stop procurement, saving time and costs. If you have any relevant intentions, please feel free to contact us for purchase at any time!
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