Nucleic acid for inhibiting thermostable polymerase and application thereof

Abstract

The invention provides nucleic acid and an application thereof, and relates to the technical field of biology, the nucleic acid provided by the invention can inhibit the activity of thermostable polymerase at normal temperature, and meanwhile, the nucleic acid has thermal startability, so that the generation of primer dimers or non-specific products at the initial stage of PCR reaction can be effectively controlled. The nucleic acid is not inactivated or degraded in the reaction process, can be folded again in the renaturation process, and is recombined to the thermostable polymerase to seal the activity of the thermostable polymerase, thereby ensuring sufficient activity of the thermostable polymerase in the system. In addition, the operation of preparing the nucleic acid modification enzyme is simple, the consumed time is short, and the batch-to-batch stability is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid for inhibiting thermostable polymerase and its application. Background technique [0002] DNA polymerase is responsible for the replication and maintenance of the genome, a central responsibility for the precise transmission of genetic information between generations. In vitro, DNA replication can be repeated many times, for example during the polymerase chain reaction. In initial studies, DNA polymerase was added at the beginning of each round of DNA replication; later, it was confirmed that thermostable DNA polymerases were available from bacteria grown at high temperatures, and that these enzymes only needed to be added once. These enzymes were not irreversibly inactivated at the high temperature of PCR. Thus, repeated cycles of the polymerase chain reaction can be performed without the need to add fresh enzyme at the beginning of each synthetic addition process. ...

AI Technical Summary

Problems solved by technology

However, because the animals themselves are exposed to other antigens in the environment during the breeding process, a certain amount of non-target antibodies will be produced, resulting in low purity of the purified antibodies; and the quality of the antibodies is also related to the health status of the animals themselves. The state will also affect the activity of the final antibody

In addition, the cell line itself will degenerate during passage and storage, which will also lead to a decrease in the activity of the antibody itself, and there are differences in antibody activity between batches, resulting in activity differences between batches of abzymes

Chemical modification of Taq DNA polymerase also has certain disadvantages: the chemical group achieves the purpose of modification by reacting with the amino acid residues in the Taq enzyme, but since the same protein contains more than one amino acid residue of the same type, and the amino acid residues The position of the base may be on the surface of the protein or inside it, leading to uncertainty in the modification

At the same time, due to the fast rate of the chemical reaction itself, it is difficult to control the entire modification process, and the chemical reaction is often accompanied by the generation of by-products, which also increases the uncertainty of the reaction.

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