What is the role of tris (Tris) in DNA extraction?
Because DNA extraction is a very pH-sensitive process, the use of tris buffer helps keep the pH stable during cell lysis and extraction. However, we need to understand the specific role of Tris buffer in DNA extraction. In this way, DNA can be better protected from pH changes during the DNA extraction process.
Tris as a buffer
Since the pH value affects and is affected by many cellular factors, maintaining a stable pH value is very important to experimental science. Tris(hydroxymethyl)aminomethane has a pKa of 8.1, which is an effective buffer with a pH between 7 and 9. Due to its neutral range, tris is a commonly used buffer in biological laboratories. However, tris buffer is sensitive to temperature and should be used at the temperature of its initial pH value to avoid inaccuracies.
Lysis of cells is the first step in DNA extraction. This is done with a buffer containing tris and EDTA (ethylenediaminetetraacetic acid). Since these ions help maintain the integrity of the cell membrane, removing them with EDTA will make the cell membrane unstable. Tris is the main buffer component; its main function is to keep the pH of the buffer at a stable point, usually 8.0. In addition, tris may interact with LPS (lipopolysaccharide) in the membrane, further destabilizing the membrane.
Tris protects DNA from pH changes
When cells divide, their DNA and contents spill into the buffer. This cell content and fragmented RNA and protein will have a great impact on the pH of the solution. Since DNA is sensitive to pH, Tris must be buffered to keep the pH at a stable point.
In the final stage of DNA extraction, the DNA itself is extracted from the solution. At this time, the DNA is soluble in the buffer. To extract from the solution, make the DNA insoluble by adding ethanol or isopropanol (isopropanol). After completion, the DNA becomes obvious in the solution as a white filamentous substance. Although DNA can be separated from the remaining cellular components in this way, it cannot be used when it is insoluble. After separation, the alcohol is removed, and the DNA must be returned to the Tris buffer before it can be used.
(Tromethamine) has always been the first among many buffers. But to really do a good job of Tris this product is not as easy as the surface. Through more than ten years of research and development and production, Desheng has produced Tris products that satisfy customers, and Desheng only needs the recognition and affirmation of customers.
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