Method for rapidly preparing gene mutation reference

Abstract

The invention discloses a method for rapidly preparing a gene mutation reference. According to the preparation method disclosed by the invention, a target required low-mutation-ratio reference can beobtained from a high-mutation-ratio reference through diluting by one step without carrying out mutation ratio verification on the reference obtained through diluting; compared with the conventional methods, the condition of firstly diluting a plurality of mutation ratio gradients and then carrying out verification one by one is also not required; and the method has the advantages of rapidness, simplicity, low verification cost and high reproducibility.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for rapidly preparing gene mutation reference products by utilizing wild-type genomic DNA and mutant DNA. Background technique [0002] In the field of cancer treatment, specific mutations carried by multiple genes are closely related to the efficacy of cancer targeted therapy drugs. Therefore, gene mutation detection has important clinical significance, and gene mutation detection technology has also been widely used. In the development and verification process of gene mutation detection technology, it is necessary to use reference products containing target gene mutations for the development and performance verification of detection reagents, which are characterized by simulating clinical samples as much as possible and containing different and known mutations Proportion. [0003] Since there are often many kinds of gene mutations to be detected, most of the m...

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Problems solved by technology

[0006] Currently, digital PCR and high-throughput sequencing are commonly used to accurately determine the proportion of target gene mutations in gene mutation reference products, and the cost of each measurement reaction is high. Therefore, when preparing mutation reference products according to conventional methods, It is necessary to prepare multiple gradients and then determine the mutation ratio one by one. This process is complex, and the number of samples for mutation ratio determination leads to a long verification process and high verification costs, which greatly restricts the preparation efficiency of mutation reference products.

If no measurement is made on the actual mutation ratio of the target gene in the diluted mutation reference products in order to save costs, the actual mutation ratio of the mutation reference products may deviate greatly from expectations, resulting in distortion of the experimental results obtained with these reference products

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