Probe and kit for detecting Top2A gene as well as detection method

A detection method and kit technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problem of low specificity of fluorescence in situ hybridization probes, long hybridization time, high signal background, etc. problem, achieve the effect of shortening hybridization time, reducing non-specific reaction, and reducing background signal

Pending Publication Date: 2019-12-31
广州简册生物技术有限公司
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Problems solved by technology

Due to the presence of some non-specific sequences in the BAC and PCR product probes, the specificity of the fluorescent in situ hybridization probes is not high, the signal background is high, and the fragments of the existing fluorescent in situ hybridization probes are usually too large, making them hybridize Low efficiency, long hybridization time

Method used

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  • Probe and kit for detecting Top2A gene as well as detection method

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Experimental program
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Effect test

Embodiment 1

[0054] FISH detection

[0055] Preparations: prepare coverslips and slides, and wipe them with 75% alcohol; preheat the metal block at 85°C; preheat the water bath at 50°C; prepare an oven at 37°C; Light wet box; prepare 150ml 2× sodium citrate buffer, 1ml 4× sodium citrate buffer, 50ml 0.4× sodium citrate buffer. Protect from light during the entire experiment.

[0056] Pretreatment of the sample to be tested: the sample to be tested is the HL60 cell line. Use PBS buffer to prepare a cell suspension. Use a pipette to absorb 10 μl of the cell suspension and drop it on the glass slide. Let it dry at room temperature until the edge of the sample is dry but the middle is still wet. When the fixative was not completely dry, 25 μl of fixative was added dropwise, and slices were obtained after the fixative dried for subsequent experiments.

[0057] Mixed probe: Top2A gene-specific probe is a mixed probe with the sequence of SEQ ID NO.1-SEQ ID NO.26. The mixed probe is dissolved in...

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Abstract

The invention provides a probe and a kit for detecting a Top2A gene as well as a detection method. The sequence of the probe provided by the invention is designed by aiming at a genome non-repetitivearea, so non-specific reaction is reduced and background signals are reduced; the probe provided by the invention is single-chain DNA, so the own pairing of the probe is reduced and the hybridizationefficiency is improved; and the probe provided by the invention is small in segment, can be rapidly combined with a targeted sequence and shortens the hybridization time.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a probe, a kit and a detection method for detecting the Top2A gene. Background technique [0002] Breast cancer is one of the most common malignant tumors in women. In recent years, the incidence rate in my country has shown a linear upward trend. Clinical studies have shown that the recurrence-free survival (Recurrence-free survival, RFS) of breast cancer patients with abnormal Top2A gene is shortened and the prognosis is poor, especially the prognosis of breast cancer patients with Top2A gene deletion is even worse. Patients with abnormal Top2A gene received anthracycline chemotherapy regimen better than patients with normal Top2A gene. Cyclophosphamide+Epirubicin+Fluorouracil (CEF) regimen can reduce the risk of recurrence by 61% and the risk of death by 51% for patients with Top2A gene amplification, while those without Top2A gene amplification Patients using the CEF program ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6834C12N15/11
CPCC12Q1/6834C12Q2565/518C12Q2563/107
Inventor 李怡赵浩
Owner 广州简册生物技术有限公司
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