METHOD FOR IDENTIFICATION OF DNA OF DOMESTIC DOG TISSUE (CANIS LUPUS FAMILIARIS) IN DRY FODDER AND MEAT SEMI-PRODUCTS Russian patent published in 2020 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology. Invention is a method for identification of DNA of domestic dog tissue (Canis lupus familiaris) in dry fodder and meat semi-products, involving DNA recovery from animal tissue by sorption method, polymerase chain reaction with fluorescent detection is carried out with 45 cycles of real-time amplification using oligonucleotide primers, probes, fluorescent dyes specific for the DNA genome region: for a specific signal for an animal and Cy5/Red for an internal control sample in form of a suspension of bacteriophage T4 with concentration of 5 × 10³ of phage particles per 1 mcl, positive control sample in the form of a mixture containing fragments of animal genomes and bacteriophage T4 with a nucleotide sequence: T4F TACATATAAATCACGCAAAGCT4R TAGTATGGCTAATCTTATTGG T4P CY5 ACATTGGCACTGACCGAGTTC, taken in ratio 1:1, and measurement of accumulation of fluorescent signals via channels of corresponding fluorescent dyes, interpretation of results based on the presence or absence of the intersection of the fluorescence curve with the threshold line, if the fluorescent signal accumulation curves extend to 35 cycles, then the result of the reaction is considered to be positive, and if the curves do not cross the threshold line or cross it after 35 cycles, the result of the reaction is negative, according to the invention, DNA is recovered from domestic dog tissue (Canis lupus familiaris) and the internal control sample is T4 bacterial phagolysate, and for the positive control sample - native T4 bacteriophage genome fragments and domestic dog tissue (Canis lupus familiaris) with the following nucleotide sequence: Canis F ATTCggCCTACATCCgTgAC forward primer Canis R AgAAgACCCCTgCTACgACT reverse primer Canis P FAM-CTTgAgTggAgTAgggCgg-BHQ1 probe, wherein for DNA of dog tissue fluorescent dye FAM/Green is used.

EFFECT: invention makes it possible to increase accuracy of species identity identification and to simplify the sampling process.

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