METHOD OF PRODUCING MODIFIED COMBINATORIAL DNA LIBRARIES BY SOLID-PHASE PRIMER EXTENSION REACTION ON PET MICROPARTICLES Russian patent published in 2020 - IPC C12Q1/68 

FIELD: organic chemistry; biochemistry; molecular biology.

SUBSTANCE: described is a method of producing combinatorial DNA libraries containing modified nucleotides for specific binding with molecular targets. Essence of invention is that combinatorial modified single-stranded DNA library is used to use a combinatorial single-strand DNA library of natural nucleotides and the complementary modified DNA is synthesized in a matrix-dependent high-specific enzymatic primer extension reaction with complete substitution of one of the natural nucleotides for the modified analogue. For primer extension reaction, Deep Vent(exo-) and KOD XL DNA polymerases and modified deoxyuridine triphosphates with C5-substituted substituents of uridine base, which are completely compatible, are proposed. In the synthesis of modified DNA one type of primers fixed on the surface of polyethylene terephthalate microparticles is used, which enables to separate and wash the carrier microparticles with the obtained fixed single-stranded modified DNA from the used DNA matrix and reaction components. Primers include groups which enable selective cleavage of the modified DNA from the carrier, without affecting its functional part by photochemical or enzymatic methods, which enables to obtain a combinatorial modified single-stranded DNA library in pure form. Besides, corresponding library is presented.

EFFECT: invention extends the methods of producing DNA libraries.

17 cl, 9 dwg, 9 ex