Method for establishing real-time fluorescent quantitative detection of PER2 promoter methylation level

• Main Authors: SUN CHENGMING, JIANG HUIHUI, SUN ZHUOKAI, LIU JIE, SUN ZHIHONG, DU JIANGDONG, YANG XIN, PENG RONGXUE
• Format: Patent
• Language: Chinese; English
• Published: 27.08.2021
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a method for establishing a real-time fluorescent quantitative detection PER2 promoter methylation level. The method comprises the following steps of S1, selecting a PER2 promoter CpG island region: 1) querying a PER2 promoter sequence according to a gene database, and 2) predicting a GpG island enrichment region contained in the PER2 promoter sequence through a CpG island online prediction website, and determining a target sequence, S2, designing a primer and a probe for PER2 promoter methylation detection according to the target sequence, S3, designing methylation and non-methylation sequences of a target sequence according to a transformation rule of bisulfite on a gene sequence, artificially synthesizing corresponding sequences, and storing the corresponding sequences in a plasmid form, and S4, establishing a real-time fluorescent quantitative PCR detection PER2 promoter methylation method, and confirming the performance of the PER2 promoter methylation method. The methylation leve