Automated reagent kit for constructing BRCA1/2 gene variation detection library

Abstract

The invention relates to an automated reagent kit for constructing a BRCA1 / 2 gene variation detection library, and provides a reagent kit for constructing an amplicon library of a BRCA1 / 2 gene. The reagent kit contains (1) a first primer for performing first-round PCR amplification; (2) a second primer for performing second-round PCR amplification; and (3) a third primer and a fourth primer, whichare used for performing third-round PCR amplification by using a second-round PCR amplification product as a formwork, and constructing an amplicon library of a target gene, wherein the first primerand the second primer are specific primers of the BRCA1 / 2 gene, and the first primer in (1), the second primer in (2) and the third primer and the fourth primer in (3) are respectively placed in separated containers. The reagent kit can be used for automation construction of the amplicon library, the tedious degree of operation is notably reduced, the demanded quantity of detection samples is notably reduced, and the whole quality of sequencing data is increased.

Description

technical field [0001] The invention relates to an automatic kit for gene detection library construction. Specifically, the present invention relates to an automated kit for constructing a BRCA1 / 2 gene variation detection library. Background technique [0002] The Human Genome Project (HGP), which cost US$3 billion and completed over 10 years, has brought earth-shaking changes to genomics research. By determining the sequence of human genome DNA, the location of genes on chromosomes is explored, and the structure of genes is clarified. And functions, interpreting all the genetic information of human beings, and human beings fully understand themselves at the molecular level for the first time. Next-generation sequencing technologies have spawned a rich and colorful genome map. [0003] Next-generation sequencing technology, also known as massively parallel sequencing (MPS), refers to the principle of "sequencing while synthesizing" to simultaneously perform parallel sequen...

AI Technical Summary

Problems solved by technology

However, this method greatly increases the complexity of the operation and the demand for detection samples.

At the same time, the current main amplicon library construction technology usually uses a multi-cycle PCR program when amplifying the target region. Due to the different amplification efficiencies of different primers, exponential PCR amplification with bidirectional primers, after multi-cycle PCR After the procedure, the uniformity of the amplified region can vary significantly, which can eventually lead to an overall poor quality of the sequencing data

[0006]For the BRCA1 / 2 gene variation detection library, the current method needs to manually add various reagents, which takes a long time to build and it is not easy to ensure the consistency of the experimental operation

In addition, as mentioned above, in the existing construction method, it is necessary to split the reaction system from one tube to multiple tubes to reduce the generation of primer dimers, and it is also necessary to consider the difference in the uniformity of the amplified region caused by the different amplification efficiencies of different primers. Significant differences and manual intervention in the PCR process, it is difficult to achieve full automation of the entire construction process

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