Multiple PCR (Polymerase Chain Reaction) primer composition, reagent and multiple PCR-breaking control system for TERT (Telomerase Reverse Transcriptase) gene whole exome next-generation sequencing

Abstract

The invention provides a multiple PCR (Polymerase Chain Reaction) primer composition, a reagent and a multiple PCR-breaking control system used for TERT (Telomerase Reverse Transcriptase) gene whole exome next-generation sequencing. The multiple PCR primer composition used for the TERT gene whole exome next-generation sequencing comprises primers disclosed from SEQ ID NO: 01 to SEQ ID NO: 32. Themultiple PCR primer composition, the reagent and the multiple PCR-breaking control system for the TERT gene whole exome next-generation sequencing solves the following existing technical problems that1: since the point mutation of the BRAF gene is mainly detected in the prior art, a detection range is limited; 2: real-time fluorescent quantitation PCR (Polymerase Chain Reaction) detection and a gene chip only can detect known sites; 3: a Sanger sequencing checkout length is smaller than a range of 800 bp; 4: low-frequency mutation can not be detected; and 5: when a plurality of sites are detected, operation is complex, and cost is high.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a multiple PCR primer composition, reagents and multiple PCR-interruption control system for second-generation sequencing of TERT gene whole exons. Background technique [0002] TERT (telomerase reverse transcriptase, telomerase reverse transcriptase) refers to the catalytic subunit with reverse transcriptase activity in telomerase, which is an important catalytic part of telomerase and affects the regulation mechanism of telomeres, thereby maintaining cell Immortalization and carcinogenesis. The TERT gene is located on chromosome 5, and its transcript NM_001193376.1 has 15 exons, with a total length of 3275 bp. TERT gene mutations are found in malignant tumors such as melanoma, lipoma, hepatoma, and glioma All have a certain detection rate, and the prognosis of positive patients is mostly poor, and they are biomarkers of related tumors. [0003] Regarding the mutation of the TERT...

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Problems solved by technology

[0004] Most of the TERT gene mutation detection in the prior art is to detect the C228T and C250T sites of the TERT gene promoter activity, which has the following limitations: 1. Real-time fluorescent quantitative PCR detection and gene chip detection cannot detect unknown sites; 2. The detection site of Sanger sequencing is limited to within 800bp; 3. When multiple sites are involved, the original technology is cumbersome and expensive; 4. It cannot detect low-frequency mutations

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