The application difference of TAE and TBE electrophoresis solution prepared by tromethamine Tris
Electrophoresis buffer is needed in the separation and purification of nucleic acid and DNA, membrane transfer, electrophoresis, and other electrophoresis experiments. The most common is the electrophoresis solution prepared with tromethamine Tris, such as TAE and TBE. What's the difference in application?
The running buffer that we usually add in DNA electrophoresis includes two buffer systems, TAE and TBE. TAE is Tris with acetic acid (acetic acid) and EDTA, and TBE is Tris with boric acid and EDTA.
TAE, TBE electrophoresis solution raw material tromethamine Tris
Which electrophoresis buffer should be used for DNA electrophoresis depends on the number of nucleic acid base pairs, electrophoresis time and other experimental conditions. Let's take a look at the different applications of TAE and TBE:
1. TAE electrophoresis solution is suitable for the separation of large DNA fragments. Fragments larger than 2kb migrate faster in TAE (in 0.8% agarose gel), and the speed is about 10% faster than in TBE .
2. TAE is more suitable for DNA fragments that need to be recovered after electrophoresis is completed.
3. For supercoiled DNA, using TAE as the electrophoresis solution has higher resolution than TBE.
4. The conductivity of TAE is lower than that of TBE. Therefore, compared with TAE, long-term electrophoresis of TBE is not likely to cause overheating in the electrophoresis tank. Therefore, it is recommended to replace TAE with TBE when long-term electrophoresis is required.
5. Boric acid contained in TBE is an enzyme inhibitor, so if you need to separate electrophoretic DNA for digestion reaction, it is recommended to use TAE as the electrophoresis buffer solution.
6. It is better to use TBE electrophoresis buffer to separate relatively small DNA fragments. For fragments less than 300bp, on a 2% agarose gel, using TBE will migrate faster than TAE.
Some electrophoresis experiments also use TPE buffer, and a buffer system composed of Tris, phosphoric acid, and EDTA. The backwash range is large, but phosphoric acid is prone to precipitation of metal ions such as Ca and Mg, which has a large impact on reactions involving enzymes, so it is better TAE and TBE are common.
Therefore, it is necessary to choose different electrophoresis buffers for DNA fragments with different base numbers and the time and purpose of electrophoresis experiments. Desheng produces the raw material tromethamine Tris used in the preparation of TAE and TBE, and can also provide ready-to-use buffers.
HEPES, as a zwitterionic buffer, increases the osmotic pressure of the cell culture system by increasing the concentration of solution ions, maintaining normal cell morphology and function, and improving cell survival rate. Widely used in cell culture, especially under specific conditions such as tumor cell culture, it is crucial to maintain cell growth and function.