METHOD FOR DETERMINING MODIFIED NUCLEOTIDES OF RNA Russian patent published in 2014 - IPC G01N33/58 

FIELD: biotechnology.

SUBSTANCE: selection of pair of oligonucleotide probes suitable for the study area of RNA, selection of a pair of donor and quencher of fluorescence with suitable optical properties is carried out. The direct chemical synthesis of modified probes is carried out, their effectiveness is assessed during melting with RNA containing known modifications. The obtained parameters of melting are assessed for each particular system. The control and the tested RNA are analysed, subjecting the mixture of probes and RNA to intense heating, slow cooling, and then the controlled heating with simultaneous detection of the fluorescence intensity, and the study parameters are selected according to the specific system. The results of melting are analysed, assessing the nature of the curves of melting, and the conclusion is made on the presence or absence of the modification in the test RNA in the presence or absence of modifications in the control RNA.

EFFECT: method is effective for detection of various types of RNA modifications and enables to detect the presence of modifications which do not prevent the formation of Watson-Crick pairs between nucleotides.

17 cl, 3 dwg, 1 ex