CDKN1A gene mutation detection specific primers and liquid chip

Abstract

The invention discloses a CDKN1A gene mutation detection liquid chip, and specific primers. The CDKN1A gene mutation detection liquid chip mainly comprises: ASPE primers, wherein each ASPE primer is composed of 5'-terminal tag sequence, and 3'-terminal specific primer sequence targeting target gene mutation sites, and the specific primer sequence comprises SEQ ID No.15 and SEQ ID No.16 targeting A7358T site, SEQ ID No.17 and SEQ ID No.18 targeting C98A site, SEQ ID No.19 and SEQ ID No.20 targeting G70A site, SEQ ID No.21 and SEQ ID No.22 targeting C258T site, SEQ ID No.23 and SEQ ID No.24 targeting T3745C site, SEQ ID No.25 and SEQ ID No.26 targeting G2763A site, and / or SEQ ID No.27 and SEQ ID No.28 targeting C4008T site; microballoons coated with different anti-tag sequences; and amplification primers. Self-agreement ratio of detection results of the liquid chip with detection results of sequencing is as high as 100%; and parallel detection of wild types and mutant types of a plurality of mutation sites is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a CDKN1A gene mutation detection specific primer and a liquid phase chip. Background technique [0002] Cyclin-dependent kinase inhibitor 1A (cyclindependent kinase inhibitor 1A, CDKN1A), also known as p21, Cip1, is located on chromosome 6 6p21.2, and the protein encoded by CDKN1A gene controls the pathway of most tumor suppressors in cells. At the same time, the protein blocks the synthesis of new strands of DNA that cancer cells need to grow and divide. The p21 gene is a member of the Clp family. It is a cyclin-dependent kinase inhibitor located downstream of the p53 gene. It can form the G1 checkpoint of the cell cycle together with p53. Because DNA damage cannot pass through without repair, it reduces Replication and accumulation of damaged DNA, thereby playing a tumor suppressor role. Studies have shown that: p21 is related to tumor d...

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Problems solved by technology

[0003] At present, CDKN1A gene mutation detection methods mainly include: Illumina fiber optic microbead chip technology, PCR-RFLP technology, fluorescence quantitative PCR technology and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), although Illumina fiber optic microbead chip technology The technology is a high-throughput detection system with high sensitivity and accuracy, but the degree of automation is low, and there are many manual operations, which are difficult to meet the needs of practical applications. The PCR-RFLP method is based on the recognition sites of restriction endonucleases caused by gene mutations. Changes, such as site loss or creation of new sites, amplify a specific fragment by PCR, and then digest the amplified product with a restriction endonuclease, and observe the size of the fragment by electrophoresis. This method is used to detect the restriction site Altered gene mutations can directly determine the genotype, but this method cannot be used for the detection of gene mutations without new restriction sites

Fluorescent quantitative PCR technology has the disadvantages of low sensitivity, easy sample contamination, and high false positive rate. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology is a soft ionization technology that has powerful and mature functions in the detection of proteins and other biological macromolecules. However, in the field of nucleic acid detection, due to the particularity of nucleic acid molecules, the detection is subject to certain restrictions.

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