Human SGIP1, SCAND3 and MYO1G gene methylation detection kit

Abstract

The invention provides a human SGIP1, SCAND3 and MYO1G gene methylation detection kit. The human SGIP1, SCAND3 and MYO1G gene methylation detection kit comprises methylation-sensitive restriction enzymes, a digestion buffer solution and a fluorescent PCR reaction solution, and the methylation-sensitive restriction enzymes are AciI, BstUI and HpaII. The detection kit adopts an enzyme mixture of themethylation-sensitive restriction enzymes being AciI, BstUI and HpaII, the digestion efficiency can be effectively improved, incomplete digestion is avoided, the false positive rate is reduced, the sensitivity is good, and the human SGIP1, SCAND3 and MYO1G gene methylation detection kit is suitable for a variety of sample types such as plasma DNA, tissue DNA and cell DNA.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a human SGIP1, SCAND3 and MYO1G gene methylation detection kit. [0002] technical background [0003] In recent years, with the development of gene detection technology, researchers have found that, in addition to gene sequence changes, gene epigenetic changes are also an important mechanism leading to the development of malignant tumors (Stefansson OA et al., The American Journal of Pathology, 2013, 183, 1052-1063). DNA methylation is one of the main forms of epigenetic changes in genes. DNA methylation refers to the process of transferring a methyl group to the 5-carbon atom of cytosine under the action of methyltransferase to form 5-methylcytosine. For DNA sequences larger than 500 bp, DNA regions with GC content greater than 55% are called CpG islands (Suzuki MM et al., Nature Reviews Genetics, 2008, 9, 465-476). 50%-60% of promoter regions in mammals are located o...

AI Technical Summary

Problems solved by technology

This technology has high sensitivity for detection of CpG island methylation and does not need to predict the sequence of the DNA to be tested. However, its disadvantages are: ① incomplete digestion may easily lead to false positive / negative results; Detection of methylation sites is also limited

However, its disadvantages are: ①The sample is easily degraded during the conversion process, resulting in impure products and decreased integrity; The specificity of sequence hybridization; ④The operation steps are cumbersome, and there are many kinds of reagents required. After the bisulfite conversion, the DNA needs to be purified and recovered before the subsequent detection steps, and the DNA purification and recovery will cause DNA loss, which further reduces the detection efficiency. sensitivity

However, its shortcomings are: ①The density of CpG sites affects the efficiency of enrichment; ②When detecting the absolute degree of methylation, it is greatly affected by the copy number

[0011] At present, there is no kit for simultaneous detection of human SGIP1, SCAND3 and MYO1G gene methylation in the market

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