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DNA helicase detection method based on graphene oxide chip

A DNA helicase, graphene oxide technology, applied in the field of biomedicine, can solve the problems of high cost and low detection throughput

Inactive Publication Date: 2015-05-13
JIANGSU UNIV
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Problems solved by technology

[0003] Most of the currently existing detection methods for DNA helicase activity pass 32 One of the labeled DNA strands of P is detected by gel electrophoresis, but this method is costly, time-consuming, and has low throughput

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  • DNA helicase detection method based on graphene oxide chip

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Embodiment Construction

[0019] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but the protection scope of the present invention is not limited thereto.

[0020] (1) Preparation of amino-modified glass slides: Add 5 ml of hydrogen peroxide (H 2 o 2 ), then

[0021] Then add 15 ml of concentrated sulfuric acid (H 2 SO 4 ), shaking slowly on a shaker or standing still for 30 min to make it

[0022] Fully react, this reaction can make the surface hydroxylated, pour off the liquid of the previous step reaction, wash 3 times with deionized water. First wash with absolute ethanol twice, then pour 20 ml of absolute ethanol, the obtained hydroxylated

[0023] The silicon wafer was transferred to a beaker and washed 3 times with absolute ethanol. Pour off the ethanol after cleaning, and quickly add a mixture of 3-aminopropyltriethoxysilane (APTES) and absolute ethanol (volume ratio 1:15), or add 15ml of absolute ethanol first,...

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Abstract

The invention belongs to protein detection in the biomedical field, and in particular provides a DNA helicase detection method based on a graphene oxide chip. The DNA helicase detection method based on the graphene oxide chip comprises the following steps: (1), preparing a slide with graphene oxide dot matrixes; (2), connecting a double-chain DNA sequence with the slide with the graphene oxide dot matrixes; (3) detecting DNA helicase, to be specific, adding different quantities of DNA helicases at different action time, performing PBS washing to scan and analyze, and detecting according to FAM fluorescence intensity change. According to the DNA helicase detection method based on the graphene oxide chip, fluorescene on FAM marked by a single-stranded DNA can be quenched by utilizing a graphene oxide; after the DNA helicase is applied to a double-stranded DNA, a single-stranded DNA separates from the complementary single-stranded DNA fixed on the graphene oxide, and the distance between the FAM and the surface of the graphene oxide is changed, so that the change of FAM fluorescence intensity is caused; by combining a high-throughput microarray chip technique, the DNA helicase can be detected with low cost and high sensitivity.

Description

technical field [0001] The invention belongs to protein detection in the field of biomedicine, in particular to a biochip-based DNA helicase detection method. Background technique [0002] DNA helicase is a protein that uses the energy provided by the hydrolysis of adenosine triphosphate (ATP) to open complementary nucleic acid double strands and obtain single strands. It plays an important role in metabolic processes such as DNA replication, repair, recombination, and transcription. They often rely on the presence of single strands and recognize the single strand structure of replication forks. Generally, it plays a role in catalyzing the unwinding of double-stranded DNA in the process of DNA replication, which plays a very important role in the reaction of virus replication and cell reproduction that requires single-stranded DNA. It has been used to treat many viral diseases, so the detection of DNA The activity of helicase is of great significance. [0003] Most of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/34
Inventor 高力时海霞李琴李娆祺周阳陈克平张春霞连超群
Owner JIANGSU UNIV
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