METHOD FOR BRCA1 AND BRCA2 GENES EXONS NUCLEOTIDE SEQUENCE DETERMINATION Russian patent published in 2017 - IPC C12N15/11 C12Q1/68 G01N33/48 

FIELD: biotechnology.

SUBSTANCE: method for BRCA1 and BRCA2 genes exons nucleotide sequence determination is described. The method includes two rounds of exon amplification with splice sites of BRCA1 and BRCA2 genes, pooled DNA sequencing after amplification by MiSeq Illumina, and data analysis. During the first round of amplification, 86 monoplex polymerase chain reactions (PCR) are performed for each DNA sample, then the amplification products are combined in equimolar amounts and are purified on magnetic particles. During the second round of amplification, the combined DNA from the first round, diluted at 1:1000, is used. At this stage, inclusion of indexing adapter nucleotide sequences takes place. Thereafter, the amplification products are normalized again and combined in equimolar amounts. Sequencing is carried out on Illumina MiSeq according to the manufacturer's instructions.

EFFECT: reduced time and cost of genes analysis, small amount of DNA required for this analysis.

2 cl, 3 tbl