LABORATORY METHOD FOR DETERMINING SEX OF BIRDS BY EXPRESSION OF RIBOCOMAL PROTEIN S6 GENE Russian patent published in 2023 - IPC C12Q1/68 A01K43/00 G01N33/53 

FIELD: molecular biology; genetic engineering.

SUBSTANCE: laboratory method for determining the sex of birds by the expression of the Ribocomal Protein S6 gene is described. Embryos of different bird species are obtained by bringing the HH36 embryo to the standard stage of development. Samples of embryonic fibroblast cells are obtained. The resulting cells are cultured in high glucose DMEM and DMEM/F12 to control the medium-related expression variation to achieve homogeneity in RNA isolation. RNA is isolated from the resulting cell cultures and used to prepare a library of sets of DNA fragments to determine the RNA sequence. The sequence of individual RNA molecules in the obtained RNA samples is determined by sequencing the prepared libraries. The quality of the resulting RNA sequence data in CAGE libraries is checked. Gene expression data is obtained from the resulting RNA samples. Based on the obtained alignments, the DNA coordinates at which individual RNA molecules are to begin to be read are determined, data on the frequency of use of certain CTSS are obtained, and gene models for the assembly are built. RNA isolated from fibroblasts is used to obtain complementary DNA. Conditions for real-time PCR are selected, a series of real-time PCR is carried out using complementary DNA from bird tissue samples as a template. The efficiency of the primers is determined in a series of DNA dilutions and the most effective primers are selected, according to the level of expression of the RPS6 gene, determined using the most effective primers used. The expression ranges of the RPS6 gene are determined for birds of different sexes in different species.

EFFECT: creation of a method for determining the sex of birds by determining the level of expression of the RPS6 gene (Ribocomal Protein S6).

1 cl, 3 dwg, 2 ex