METHOD OF MAPPING POSITIONS OF ROW OF METHYLATED NUCLEOTIDE SEQUENCES Pu(5mC)GPy IN EXTENDED DNA FOR CONSTRUCTING EPIGENETIC PROFILE AND DETECTION OF ABNORMAL METHYLATED DNA SECTIONS Russian patent published in 2016 - IPC C12N15/10 C12Q1/68 G06F19/18 

FIELD: biotechnology.

SUBSTANCE: method envisages mapping positions of methylated nucleotide sequences Pu(5mC)GPy row in extended DNA for constructing epigenetic profile and detection of abnormal methylated DNA sections. Method involves extraction of highly pure genomic DNA, which is hydrolysed in reaction buffer of methyl dependent site-specific endonuclease GlaI during period of time and at temperature sufficient for complete product hydrolysis to obtain mixture of genomic DNA fragments. Said DNA fragments mixture is extracted from solution of deposition, and for further analysis of extracted GlaI-DNA fragments with size of 150-500 bps, most optimum suitable for NGS-sequence analysis. Determined nucleotide composition of genomic DNA formed short GlaI ends fragments by NGS-sequencing, software filtering obtained data in order to exclude fragments with low-quality reading, with ends non-specific for GlaI fragments, and based on presence of GPy dinucleotide on 5′-ends of sequences and determining positions in nucleotide sequences Pu(5mC)GPy reference genome of filtered GlaI fragments using any software of “genomic assembler” class.

EFFECT: implementation of method allows simplifying, improving reliability and validity of large number of methylated PuCGPy sites in the genome method of positions (mapping) determining.

5 cl, 3 dwg