Cytosine methylation excavation method

Abstract

The invention discloses a cytosine methylation excavation method which comprises the following steps: A) acquisition of target data: amplifying gene groups from a same source after an HpaII enzyme digestion and an MspI enzyme digestion are carried out on the gene groups by methylation sensitive enzymes and performing high-throughput sequencing on the gene groups after the enzyme digestions; B) establishment of bases with an AFSM (amplification finite-state machine) technology: establishing DNA libraries of the HpaII enzyme digestion and an MPaI enzyme digestion for a same sample and respectively adding barcode linker sequences; C) interpretation of enzyme digestion sites according to original data of the sequencing; D) output of a bam comparison file by performing map analysis and assembly on the marked original data; E) methylation data excavation of the bam comparison file. The method is a first technology of performing methylation typing on cytosine methylation data after the high-throughput sequencing and the enzyme digestions of the methylation sensitive enzymes and identifying the abundance of methylation sites; a simple and efficient solution with low cost is provided for scientific research personnel to excavate methylation information.

Description

technical field [0001] The invention relates to the technical field of biological information, in particular to a method and system for methylation mining of high-throughput sequencing sequences. Background technique [0002] Sodium bisulfite sequencing (bisulfite genomic sequencing) is a method based on MSP to further study the methylation of each site of CpG island. Bisulfite deaminates the unmethylated cytosine in DNA into uracil, while the methylated cytosine remains unchanged for PCR amplification (try to avoid CpG in primer design to avoid methyl Influenced by chemical factors) to the desired fragment, all uracil is converted into thymine. Finally, the PCR product is sequenced and compared with the unprocessed sequence to determine whether the CpG site is methylated. Although this method is a method with high reliability and accuracy, it can clarify the methylation status of each CpG site in the target fragment. In searching for meaningful key CpG sites, there are...

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Problems solved by technology

Its disadvantage is that it takes too much time and money. At least 10 clones must be sequenced to obtain reliable data. A large number of clones and plasmid extraction and sequencing are required. The process is cumbersome and expensive.

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