Application of DTT |CAS:3483-12-3|in Nucleic Acid Extraction

DTT is a small molecule organic reducing agent. DTT is mainly used in nucleic acid extraction/protection, IVD kits, reduction of disulfide bonds in proteins, electroless nickel plating, etc. The editors of YACOO will introduce the application of DTT in nucleic acid extraction.

Nucleic acid can be divided into DNA and RNA. Nucleic acid can exist stably in vivo, but it is easy to cause nucleic acid degradation due to the release of nuclease after ex vivo. With the advancement of medical technology, the application of molecular disease diagnosis is increasing, and the demand for nucleic acid extraction and preservation is also increasing. Therefore, it is particularly critical to ensure the stability of clinical samples. At present, the commonly used method is to use high concentration of super-disordered salts, such as guanidine isothiocyanate, urea, etc., as stabilizers for nucleases. Guanidine salts, as toxic chemicals, especially guanidine hydrochloride and guanidine isothiocyanate, are produced by few manufacturers. Therefore, it is very necessary to invent a nucleic acid protection solution for biological samples that does not contain guanidine salts or has a low content of guanidine salts.

CN112280775A Provides a nucleic acid protection solution, which is specifically composed of 5 parts: nuclease inhibitor (urea or guanidine isothiocyanate), surfactant (sodium lauryl sarcosinate, 3-Sulfopropyltetradecyldimethylbetaine or Triton X-100), Penetrating Agents (Ammonium Sulfate, Ammonium Chloride or Aluminum Ammonium Sulfate), Chelating Agents (Sodium Citrate, Disodium EDTA, EDTA) dipotassium), reducing agents (DTT, sodium citrate, disodium EDTA, dipotassium EDTA).

The preferred solution of the nucleic acid protection solution in the patent is: urea 60g/L, guanidine isothiocyanate 60g/L, Triton X-100 3g/L, EDTA 6g/L, ammonium sulfate 70g/L, aluminum sulfate Ammonium 30g/L, DTT 5g/L, sodium citrate 5g/L and balance water.

This patent can prevent the degradation of nucleic acid in the sample through the synergistic effect of each component. The nucleic acid protection solution of the sample contains a small amount or no guanidine salt, which not only reduces the toxicity of the protection solution, but also avoids the problem of affecting production due to the low yield of guanidine salts on the market.

References

CN112280775A A kind of biological sample nucleic acid protection solution and application