DNA probe library for hybridization with MSI (micro-satellite instability) related micro-satellite sites, detection method and kit

A technology of microsatellite sites and DNA probes, which is applied in the direction of DNA preparation, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of high sample quality requirements, low throughput, and increased detection costs The effect of detection cycle, sensitivity improvement, and accuracy assurance

A technology of microsatellite sites and DNA probes, which is applied in the direction of DNA preparation, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of high sample quality requirements, low throughput, and increased detection costs The effect of detection cycle, sensitivity improvement, and accuracy assurance

CN107475375AActive Publication Date: 2017-12-15GENESEEQ TECH INC
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  • DNA probe library for hybridization with MSI (micro-satellite instability) related micro-satellite sites, detection method and kit
  • DNA probe library for hybridization with MSI (micro-satellite instability) related micro-satellite sites, detection method and kit
  • DNA probe library for hybridization with MSI (micro-satellite instability) related micro-satellite sites, detection method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0144] Example 1: Enrichment and detection of MSI-related genes

[0145] 1. Prepare the DNA sample bank to be tested

[0146] 1. Extract ctDNA from diseased plasma samples and then fragment it.

[0147] 1.1 DNA extraction

[0148] After the clinical blood sample of a patient with colon cancer was collected, immediately at 2700xg for 10 minutes, the upper serum was collected in a clean tube, stored at -80°C for later use, and extracted with the QIAGEN Neasy Blood & Tissue Kit (QIAGEN, Hilden, Germany) Peripheral blood DNA, QIAamp Circulating Nucleic Acid Kit to extract circulating tumor DNA. Operate according to the instructions in the manual.

[0149] Use spectrophotometer and gel electrophoresis system to detect the quality and concentration of DNA. The 260nm absorbance of DNA is greater than 0.05, and the ratio of absorbance A260 / A280 is between 1.8 and 2 is qualified.

[0150] 1.1 DNA fragmentation

[0151] Dilute 3 µg of high-quality genomic DNA to 120 µl with low TE...

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Abstract

The invention provides a DNA probe library for hybridization with MSI (micro-satellite instability) detection related micro-satellite sites. The DNA probe library comprises one or more DNA probes capable of hybridizing with the MSI state related microsatellite sites. In the DNA probes, the MSI related micro-satellite sites are shown in the following sequences: SEQ ID NO. 1-66. Besides, the invention provides a method for enriching and detecting the MSI related micro-satellite sites by the probe library. With combination of the method with the next generation sequencing technology (NGS), sensitivity, accuracy and comprehensiveness of MSI detection can be greatly improved.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a method for enrichment and detection of microsatellite sites related to microsatellite instability (MSI). This method can accurately enrich the specific fragments of MSI-related microsatellite loci, and the obtained DNA sample library can be further combined with next-generation sequencing technology (NGS), and quantitatively evaluate the patient's MSI status through bioinformatics analysis, providing a basis for the diagnosis of tumors. Prognosis, and the design of clinical treatment plan to provide guidance and theoretical basis. Background technique [0002] Microsatellite instability (Micro-Satellite Instability, MSI) refers to the change in the length of microsatellite alleles due to abnormal insertion or removal of repetitive sequences during DNA replication, and the change is due to various reasons (such as DNA mismatch repair The mechanism of gene promoter methylation in...

Claims

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Application Information

Patent Timeline
15 Dec 2017
Publication
CN107475375A
IPC
C12Q1/68; C40B40/06
CPC
C12Q1/6869; C12Q1/6886; C40B40/06; C12Q2600/118; C12Q2600/156; C12Q2535/122; C12N15/1093; C12Q1/6806
Inventors
邵阳; 蒋斯明