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Probe library, detection method and kit for detecting effectiveness of DNA base excision repair pathways

A kit and effective technology, applied in the field of probe libraries, detection methods and kits for detecting the effectiveness of DNA base excision repair pathways, can solve problems such as increasing tumor risk, increasing tumor ratio, and decreasing activity

Inactive Publication Date: 2020-02-14
SUZHOU HEALTH COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the OGG1 gene R46Q, S326C and other variants have reduced enzyme activity, and the probability of suffering from kidney cancer and lung cancer increases; the XRCC1 gene R194W variant enhances the efficiency of the DNA base excision repair pathway and enhances the ability to resist tumorigenesis, while XRCC1 The gene R399Q variant leads to a decrease in the efficiency of the DNA base excision repair pathway, which increases the risk of cancer; the PARP1 gene V762A variant enzyme activity decreases, resulting in an increase in the rate of various tumors; in addition, DNA polymerase Polβ at about Mutations occur in 30% of tumors, including frameshift mutations, fragment deletions, and point mutations

Method used

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  • Probe library, detection method and kit for detecting effectiveness of DNA base excision repair pathways
  • Probe library, detection method and kit for detecting effectiveness of DNA base excision repair pathways
  • Probe library, detection method and kit for detecting effectiveness of DNA base excision repair pathways

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Experimental program
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Effect test

Embodiment 1

[0117] Example 1 Detection of mutations in APE1, PARP1, OGG1, FEN1 genes

[0118] 1. Build a sample library

[0119] 1. Extraction of DNA

[0120] According to the routine DNA extraction method of tissue samples, commercial kits were used to extract sample DNA.

[0121] 2. DNA Fragmentation

[0122] Fragment the DNA sample using a DNA shredder to make the fragment length 150-200bp.

[0123] 3. DNA sample library quality inspection

[0124] Use a bioanalyzer to perform qualitative and quantitative DNA analysis to confirm that the peak length of the DNA fragment is reasonable.

[0125] 4. DNA end repair

[0126] Use T4 polymerase and Klenow E. coli polymerase fragments to blunt the DNA 5' overhanging sticky ends and 3' overhanging sticky ends to generate blunt ends for subsequent blunt end ligation. The reaction system is shown in Table 1, and the reaction was carried out in a PCR amplification instrument at 20° C. for 30 min.

[0127] 5. Add base A to the 3' end of the D...

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Abstract

The invention provides a probe library, detection method and kit for detecting effectiveness of DNA base excision repair pathways. The probe library consists of probes shown in SEQ ID NO.1-236, and the detection kit prepared by using the probe library can be used for detecting genes related to the DNA base excision repair pathways. To-be-analyzed target DNA fragments are enriched by the probe library, and then, the enriched DNA fragments are sequenced by a sequencing means to realize effectiveness for gene detection. The genes related to the DNA base excision repair pathways can be used for predicting tendency of tumorigenesis, evaluating malignant degree of tumor and clinical prognosis and guiding screening, research and development of new drugs, therefore, the probe library, detection method and kit for detecting effectiveness of DNA base excision repair pathways have important guidance significance in the research direction of gene functionality.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a probe library, a detection method and a kit for detecting the effectiveness of a DNA base excision repair pathway. Background technique [0002] Abnormal DNA damage repair pathways are closely related to the tendency of tumorigenesis and the sensitivity of DNA damage-induced anti-tumor drugs to cells. The DNA base excision repair pathway mainly repairs DNA bases damaged by endogenous oxygen free radicals, as well as DNA damage caused by alkylating antitumor drugs such as TMZ, and is one of the most important pathways in the DNA repair mechanism. The repair process is specifically divided into two pathways: single-nucleotide excision repair (Single-nucleotide BER, SN-BER) and long fragment excision repair (Long-patch BER, LP-BER). When the base is damaged, both pathways start with the recognition of the damage by DNA glycosylase, and then cut the glycosidic b...

Claims

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Application Information

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IPC IPC(8): C12Q1/6827C12N15/11
CPCC12Q1/6827
Inventor 刘松柏杜佳慧
Owner SUZHOU HEALTH COLLEGE