In-situ gene mutation targeted detection method based on high-density space dot matrix

Abstract

The invention discloses an in-situ gene mutation targeted detection method based on a high-density space dot matrix. The in-situ gene mutation targeted detection method specifically comprises the following steps: capturing specific information of RNA (particularly mRNA related to protein coding) through a high-density dot matrix technology; the captured RNA is subjected to reverse transcription to form full-length DNA, the full-length DNA is used as a template, amplification is carried out through a universal primer designed in advance, and linker sequences and cohesive ends designed in advance are introduced to the fifth end and the third end; the DNA products can be connected through the T4 ligase to form cyclized DNA; the cyclized DNA is used as a template, amplification is performed through primers designed for different DNA mutation sites, and high-throughput sequencing linker sequences designed in advance are introduced into the fifth end and the third end of a product; and completing the construction of a high-throughput sequencing library, and sequencing on a machine. The method provided by the invention aims at pathological tissue slices, and RNA and DNA capture of the sample can be completed at the same time.

Description

technical field [0001] The invention relates to the technical field of gene mutation detection, in particular to an in-situ gene mutation targeted detection method based on a high-density spatial lattice. Background technique [0002] The treatment of many diseases in life involves the genetic level, such as the treatment of cancer. Most of these diseases are treated with targeted therapy. Targeted therapy is a treatment method at the cellular and molecular level that targets a well-defined cancer-causing site (the site can be a protein molecule inside a tumor cell or a gene segment). Corresponding therapeutic drugs can be designed. When the drugs enter the body, they will specifically select carcinogenic sites to combine and act, causing the tumor to die specifically without affecting the normal tissue cells around the tumor. Therefore, molecular targeted therapy is also known as "Bio Missile". Patients need targeted detection to determine gene mutation before targeted t...

AI Technical Summary

Problems solved by technology

Patients need to undergo targeted detection to determine the gene mutation before undergoing targeted therapy, but the existing targeted detection method needs to first determine the pathological condition in the tissue section, and then use other methods to detect the gene mutation information. separate operations