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Fluorescent quantitative PCR (polymerase chain reaction) detection kit for TOP2A (topoisomerase2A) gene expression quantity and application of kit

A technology of gene expression and fluorescence quantification, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of unclear mode of action of non-embedded inhibitors, and achieve high sensitivity, good specificity, good The effect of applying the foreground

Pending Publication Date: 2016-01-20
SUZHOU BEISIMAI MEDICAL INSTR CO LTD
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

Mode of action of non-intercalated inhibitors is unclear

Method used

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  • Fluorescent quantitative PCR (polymerase chain reaction) detection kit for TOP2A (topoisomerase2A) gene expression quantity and application of kit
  • Fluorescent quantitative PCR (polymerase chain reaction) detection kit for TOP2A (topoisomerase2A) gene expression quantity and application of kit
  • Fluorescent quantitative PCR (polymerase chain reaction) detection kit for TOP2A (topoisomerase2A) gene expression quantity and application of kit

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Experimental program
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Effect test

Embodiment 1

[0027] (1) Sample preparation: take surgically removed tissue from the patient, and extract RNA (concentration not less than 100ng, A260 / 280 not less than 1.9).

[0028] (2) Sample treatment: Take 100ng of RNA, add 5 μl of reverse transcription reagent (promega kit), add double distilled water to 20 μl, and perform reverse transcription reaction according to the following conditions: 15 minutes at 25°C; 1 hour at 50°C; 85°C 5 minutes at °C; 5 minutes at 4°C.

[0029] (3) Fluorescent quantitative PCR reaction:

[0030] Use the reverse transcription product in (2) as a template: Take 5 μl of the above reverse transcription template and add it to a 0.1 ml 8-strip tube (containing 15 μl of fluorescent quantitative PCR reagent). At the same time in the standard

[0031] Add standard products with corresponding gradient concentrations to the quasi-tube (containing 15 μl of fluorescence quantitative PCR reaction reagent); add 5 μl of the above cDNA template to the internal referenc...

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Abstract

The invention relates to a fluorescent quantitative PCR (polymerase chain reaction) kit for detecting human TOP2A (topoisomerase2A) gene mutation. The kit mainly comprises specific primers and a PCR reaction reagent, wherein sequences of the specific primers are as follows: the sequence of an upstream primer is represented as 5'-GGGAGAGTGATGACTTCCATATGGA-3', and the sequence of a downstream primer is represented as 5'-AACACCTTCCCCAAACTAAATTCAG-3'. The kit has the main benefits as follows: the kit is used for detecting the human TOP2A gene expression quantity, is good in specificity and high in sensitivity and has the better application prospect.

Description

(1) Technical field [0001] The invention relates to a human topoisomerism IIα (TOP2A) gene fluorescent quantitative PCR detection kit and application thereof. (2) Background technology [0002] Topoisomerase (TOP) is a class of enzymes present in the nucleus, which can catalyze the breakage and combination of DNA strands, thereby inhibiting the topological state of DNA. There are two main types of topoisomerases in mammals, and topoisomerase I catalyzes a change in the topoisomeric state of DNA replication by forming transient single-strand cleavage binding loops. In contrast, topoisomerase II causes transient cleavage of double-strand enzyme bridges, followed by opening and reclosing, to change the topological state of DNA. Topoisomerase II includes two isoenzymes, TOPIIa and TOPIIβ. The coding gene of TOPIIa is TOP2A, located at 17ql2.21. TOP2A is most abundant in rapidly proliferating cells, and its expression is limited to the cell cycle from S phase to G2 / M phase. T...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 刘小龙张云杨福太高运臻李雨阳
Owner SUZHOU BEISIMAI MEDICAL INSTR CO LTD
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