DNA polymerase immobilization method and application thereof

Abstract

The invention belongs to the field of enzyme engineering and particularly relates to a DNA polymerase immobilization method and an application thereof. The immobilization method comprises steps as follows: a modified chitosan water solution is prepared firstly, an adsorption reaction solution of DNA polymerase is prepared, a crosslinking reaction solution of the DNA polymerase is prepared, the crosslinking reaction solution and wood pulp sponge are mixed, a carrageenin water solution is added, the mixture is centrifuged and washed and finally freeze-dried, and the immobilized DNA polymerase isobtained, and the prepared immobilized DNA polymerase has higher activity and better thermal stability; the prepared immobilized DNA polymerase has the highest activity and the best thermal stabilityby selecting a specific protein flocculant, specific adsorption reaction conditions, specific crosslinking reaction conditions, a specific crosslinking agent, a specific buffer solution, specific carriers, a specific PCR accelerator and specific freeze-drying conditions.

Description

technical field [0001] The invention belongs to the field of enzyme engineering, and in particular relates to a DNA polymerase immobilization method and application thereof. Background technique [0002] Polymerase chain reaction (PCR) refers to: the DNA will be denatured into a single strand at a high temperature of 95°C in vitro, and the single strand will combine with the primer according to the principle of complementary base pairing at a low temperature of about 60°C. When the optimum reaction temperature of DNA polymerase is around 72°C, DNA polymerase will synthesize complementary chain properties along the direction from phosphoric acid to five-carbon sugar (5'-3'), and realize the molecular biology technology of amplifying and amplifying specific DNA fragments . [0003] Taq DNA polymerase is the heat-resistant DNA polymerase with the highest amplification efficiency. It can amplify DNA fragments below 6kb very well, and the amplification base error rate is 10 -5 ...

AI Technical Summary

Problems solved by technology

[0007] For this reason, the first technical problem to be solved by the present invention is the problem that the activity of the immobilized TaqDNA polymerase prepared by the prior art is low, and the second technical problem to be solved by the present invention is the immobilized TaqDNA polymerase prepared by the prior art. In order to solve the problem of poor stability of Taq DNA polymerase, an immobilization method of DNA polymerase is proposed, and then the application of the immobilized DNA polymerase prepared by it is provided.