An improved buffer system applied to pcr or rt-pcr and its application

Abstract

The invention discloses an improved buffer system applied to PCR or RT-PCR. The improved buffer system is formed by mixing 3-(N-morpholine) propanesulfonic acid, Tris and trisodium citrate, wherein a solution state is alkalinity; the molar ratio of the 3-(N-morpholine) propanesulfonic acid to the Tris to the trisodium citrate is (20-100):(10-50):(1-5); and the pH of the improved buffer system is 8.3-9. The improved buffer system has higher buffer stability, high amplification efficiency and good detection sensitivity and precision.

Description

technical field [0001] The invention relates to the field of molecular detection, and provides an improved buffer system applied to PCR or RT-PCR and its application. Background technique [0002] At present, PCR technology is widely used in biology, medical treatment, food and other fields. Providing a stable PCR or RT-PCR buffer system is a key factor for efficient performance of various enzymes in the PCR process. The Tris-HCl buffer system is the most classic PCR reaction system, which ensures the stability of pH in PCR work and other functions. However, Tris-HCl buffer is a dual-polarized ionic buffer with a pKa of 8.3 (20°C) and a △pKa of 0.021 / °C. It is relatively temperature sensitive, and the pH decreases as the temperature increases. Under typical thermocycling conditions, when the extension temperature is 72°C, the real pH value is between 6.8-7.8, and the buffering capacity is extremely poor at pH less than 7.5. For many hot-start enzymes, pH changes lead to d...

AI Technical Summary

Problems solved by technology

[0004] Although the above scheme has solved the deficiencies of the prior art to a certain extent, the overall design is not reasonable enough.

At present, there is still a lack of an improved PCR or RT-PCR buffer system and its application with good detection sensitivity.

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