Whole genome overall 5hmC detection method and application thereof
A technology of genome and detection reagents, applied in the fields of biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, etc., can solve the problems of unsatisfactory detection conditions, increase the difficulty of detection, and high sample requirements, and achieve the reduction of sample volume, The effect of improving sensitivity and high sensitivity
Active Publication Date: 2021-11-12
SUN YAT SEN UNIV
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However, due to the extremely low content of 5hmC in the genome, and its structure is highly similar to other bases, coupled with the complexity and sensitivity limitations of various detection instruments, the actual detection of 5hmC in the genome is not ideal.
Moreover, the amount of sample to detect 5hmC usually needs to reach the microgram level, and the requirements for the sample are relatively high, which further increases the difficulty of its detection
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[0062] In order to make the object, technical solution and technical effect of the present invention clearer, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the specific implementations described in this specification are only for explaining the present invention, not for limiting the present invention.
[0063] The experimental materials and reagents used, unless otherwise specified, are conventional consumables and reagents that can be obtained from commercial channels.
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Abstract
The invention discloses a whole genome overall 5hmC detection method and application thereof. According to the whole genome overall 5hmC detection method, PolyT stable fluorescent copper nanoparticles are used for detecting 5hmC, ddTTP and TdT are added for end capping, UDP-6-N3-Glu and T4-beta GT are added after TdT is heated and inactivated, a probe and the copper nanoparticles are added, molecules of a large number of fluorescent signals are modified on a 5hmC site, efficient 5hmC signal amplification is achieved, the fluorescence detection sensitivity is improved, the sample size can reach 5ng, no large instrument or magnetic bead enrichment is needed, and the method can be suitable for detecting all nucleic acids.
Description
technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a whole genome 5hmC detection method and an application thereof. Background technique [0002] DNA methylation is an important research content of epigenetics. Cytosine in the genome, as part of the genetic code, carries epigenetic information through the chemical modification of its pyrimidine ring, and these chemical modifications are mainly 5-methylcytosine (5mC) and its oxidation product 5-hydroxymethylcytosine ( 5hmC), 5-formylcytosine (5fC) and 5-carboxycytosine (5caC). 5mC can silence gene expression, but it can re-express silenced genes through TET enzyme-mediated oxidation and a series of active demethylation processes. Therefore, the oxidation products of 5mC are of great significance for precise gene regulation. Moreover, 5hmC is also a potential cancer marker, and abnormal 5hmC levels are closely related to the occurrence and development of cancer. Therefor...
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IPC IPC(8): C12Q1/6886C12Q1/6851C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q1/6858C12Q2600/154C12Q2525/173C12Q2563/107C12Q2563/155C12Q2565/125C12Q2545/113Y02A50/30
Inventor 戴宗于振宁邹小勇
Owner SUN YAT SEN UNIV
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