Capture library and kit for detection of 50 tumor-associated hotspot mutation genes

Abstract

The invention discloses a kit and a method for detecting 50 tumor-relevant hotspot mutation genes. According to the method, a capture method of 207 hotspot mutation regions of 50 relevant genes (including oncogenes and cancer suppressor genes) and a preparation method of a liquid-phase hybridization library used in capturing are researched aiming at oncology. The method particularly comprises the steps of selecting sequence information of 207 hotspot mutation regions, designing a PCR primer, preparing a biotin labeled liquid-phase hybridization library by virtue of a PCR method, hybridizing the biotin labeled liquid-phase hybridization library with a constructed target genome DNA library, and carrying out high-throughput sequencing and bioinformatics analysis on captured fragments, so as to obtain the mutation condition of nucleic acid sequence sites of a sample. By virtue of the method provided by the invention, high-quality sequencing data can be obtained, the relatively high capturing efficiency and the relatively good uniform capturing of the target sites can be realized, the disadvantages of low efficiency, poor uniformity and the like of solid phase hybridization are solved, and the sequencing cost is greatly lowered.

Description

technical field

[0001] The invention belongs to the technical field of gene detection, and relates to a kit and a detection method for detecting cancer-related genes, in particular to a preparation method and technology of a hybridized sequence capture library. Background technique

[0002] Malignant tumors, commonly known as cancer, according to statistics, cancer has become the leading cause of death of urban and rural residents in my country. In the past hundred years, people have been studying cancer without interruption, but the specific etiology is still uncertain. With the development of science, scientists have discovered that oncogenes and tumor suppressor genes play a key role in the occurrence of cancer. An oncogene can generally be defined as a gene whose abnormal expression or abnormal expression product directly determines the generation of malignant phenotype of cells. It is now known that in tumorigenesis, viruses, chemical carcinogens and radiation as envi...

Examples

Embodiment 1

[0036] Specifically, on the basis of the present invention based on the existing or obtained information on 50 hot-spot mutation genes related to tumors, a detection kit for 207 hypermutated regions of 50 hot-spot mutation genes related to tumors was constructed. The kit first The extracted sample DNA is ultrasonically fragmented into 200-250bp DNA fragments (but not limited to the ultrasonic fragmentation method), and the DNA fragments are connected to the index adapter through end repair, adding "A" and ligation, and then use the constructed The capture library is hybridized with the DNA fragments containing the target gene region to be captured, and the DNA fragments of the tumor-associated hotspot mutation genes are captured. These DNA fragments are sequenced, and the sample detection results are obtained after bioinformatics data analysis. The technical solution includes the entire experimental process from sample genomic DNA to the output of sequencing results, mainly inc...

technical field

[0001] The invention belongs to the technical field of gene detection, and relates to a kit and a detection method for detecting cancer-related genes, in particular to a preparation method and technology of a hybridized sequence capture library. Background technique

[0002] Malignant tumors, commonly known as cancer, according to statistics, cancer has become the leading cause of death of urban and rural residents in my country. In the past hundred years, people have been studying cancer without interruption, but the specific etiology is still uncertain. With the development of science, scientists have discovered that oncogenes and tumor suppressor genes play a key role in the occurrence of cancer. An oncogene can generally be defined as a gene whose abnormal expression or abnormal expression product directly determines the generation of malignant phenotype of cells. It is now known that in tumorigenesis, viruses, chemical carcinogens and radiation as envi...

Examples

Embodiment 1

[0036] Specifically, on the basis of the present invention based on the existing or obtained information on 50 hot-spot mutation genes related to tumors, a detection kit for 207 hypermutated regions of 50 hot-spot mutation genes related to tumors was constructed. The kit first The extracted sample DNA is ultrasonically fragmented into 200-250bp DNA fragments (but not limited to the ultrasonic fragmentation method), and the DNA fragments are connected to the index adapter through end repair, adding "A" and ligation, and then use the constructed The capture library is hybridized with the DNA fragments containing the target gene region to be captured, and the DNA fragments of the tumor-associated hotspot mutation genes are captured. These DNA fragments are sequenced, and the sample detection results are obtained after bioinformatics data analysis. The technical solution includes the entire experimental process from sample genomic DNA to the output of sequencing results, mainly inc...

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