Process for quickly detecting DNA methylation

Abstract

The invention relates to the medical biology detection technical field and discloses a method for quickly detecting DNA methylation. Aiming at the disadvantages of the prior method, the method provided by the invention improves the following three aspects: 1. catalyst TAC and guanidine hydrochloride are added to accelerate the modification reaction; 2. short-time high-temperature melting is performed during the modification reaction for times so as to fully expose basic groups to transform cytimidine which is not made from methylation modification to sulfonated uracil; 3. aiming at fussy dialysis desalination steps after the modification, when a modified DNA product is purified, the modified DNA product is combined into a glass fiber membrane or pellosil or other pillars for washing desalination, then is treated by sulfated reagent and eluted, thereby obtaining a methylated PCR template which can be directly used for PCR augmentation and greatly saves desalination time. The method has low cost, short detection time and good repetitiveness.

Description

technical field [0001] The invention relates to the technical field of medical biological detection, in particular to a method for rapidly detecting DNA methylation. Background technique [0002] Abnormal methylation of genes is closely related to the occurrence and development of tumors. In vertebrates, CpG dinucleotides are the main sites of DNA methylation. CpG often exists in clusters, and a section of DNA rich in CpG in the genome is called CpG island (CpGisland). CpG islands are often located near transcriptional regulatory regions, and the study of DNA methylation is inseparable from the study of CpG islands. Abnormal methylation has appeared in early cancer, precancerous lesions, and even some benign lesions. Hypermethylation of CpG islands in the promoter regions of multiple tumor suppressor genes has been found in precancerous lesions of many malignant tumors. According to reports, abnormal methylation of multiple genes related to pancreatic cancer has been detec...

AI Technical Summary

Problems solved by technology

[0039] This traditional detection method has the following disadvantages: 1. The operation is cumbersome, especially in the process of sodium bisulfite modification, the modification reaction is slow and takes a long time, usually 16-18 hours; 2. After the DNA is denatured into a single strand, the single The bases on the chain will also pair with each other, which may cause insufficient base exposure, resulting in incomplete sodium bisulfite modification, low experimental repeatability, and affecting the reliability of the results; 3. After sodium bisulfite modification, The desalting time of dialysis is also very long and the workload is heavy, which restricts the research of MS-PCR

However, if the DNA methylation detection kit (EZ DNA Methylation-Gold TM Kit), although the operation is convenient and the detection time is short, but the price is expensive, and the PCR template obtained each time can only be used for the methylation detection of 2 genes, which limits the multi-gene detection and cannot meet the needs of most experiments.

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