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Methylation
Experimental principle
DNA methylation exists in most eukaryotes, and generally occurs in the 5'UTR region of cytosine in CpG dinucleotides, which plays a regulatory role. About 80% of the cytosine of CpG is methylated, and 20% is in the CpG island of the gene regulatory region. The principle of BSP methylation sequencing (Bisulfite Genomic Sequencing PCR) is through bisulfite treatment of genomic DNA. Unmethylated cytosine is deaminated and converted into uracil. In the subsequent PCR reaction, uracil is converted into Thymine, and methylated cytosine cannot be deaminated and is retained when the reaction is completed. We will perform TA cloning and sequencing on the amplified PCR products to analyze the specific conditions of methylation. This method has high accuracy and is the gold standard for methylation detection.
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