Primer group and kit for detecting by combining breaking point crossing PCR (Polymerase Chain Reaction) with multiple PCR (Polymerase Chain Reaction)

Abstract

The invention relates to the technical field of sequencing, in particular to a primer group and a kit for gene detection through combination of breaking point crossing PCR and multiple PCR, and provides a primer group for sequencing through combination of breaking point crossing PCR and multiple PCR, the primer group comprises a first primer, a second primer, a third primer, a fourth primer and a fifth primer, the first primer sequentially comprises a specific primer and a first sequencing tag from the 5'end to the 3 'end, the second primer sequentially comprises a specific primer and a second sequencing tag from the 5' end to the 3 'end, and the specific primers comprise a forward primer of an upper breaking point, a reverse primer of the upper breaking point, a forward primer of a lower breaking point and a reverse primer of the lower breaking point. By adopting the primer group, whether gene large-area deletion or insertion exists in the whole exon of a sample gene can be quickly analyzed, the speed is high, and the accuracy rate is high.

Description

technical field [0001] The present application belongs to the technical field of sequencing, and in particular relates to a primer set and a kit for detecting large-area deletion or insertion of genes by cross-breakpoint PCR combined with multiplex PCR. Background technique [0002] Inherited metabolic disease is a single gene disease. Due to gene mutation, the function of synthetic enzymes, receptors, carriers and other proteins is defective, resulting in various abnormalities in the synthesis, metabolism, and transport of biochemical substances in the body, resulting in a series of clinical symptoms. class diseases. A small number of these diseases are currently treatable and preventable, and the development of dietary therapy and drug therapy will improve the prognosis of some patients. [0003] The application of high-throughput sequencing technology has been more than 10 years, and its high-throughput, high-sensitivity and low-cost advantages make it have broad clinica...

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Problems solved by technology

[0008] The purpose of this application is to provide a primer set and kit for sequencing by combining multiplex PCR across breakpoints, aiming to solve the problem that some large fragment insertions or deletions spanning exons cannot be detected by sequencing detection in the prior art. The problem with the accuracy of test results

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