HIGH THROUGH-PUT ANALYSIS OF TRANSGENE BORDERS Russian patent published in 2016 - IPC C12Q1/68 C12P19/34 

FIELD: biochemistry.

SUBSTANCE: present invention relates to biochemistry, in particular, to a method of determining and sequencing unknown DNA sequences, which flank known sequences in isolated DNA. Method is realised by first performing hydrolysis of isolated DNA, containing a part or all of known polynucleotide sequence and adjoining unknown polynucleotide sequence, with suitable restriction enzymes. Further, method includes synthesizing a complementary strand by hydrolysed polynucleotide restriction fragments using oligonucleotide sequence of a primer, having a chemical binding structure, bound with 5′ end of oligonucleotide sequence of primer. Then said complementary strand is isolated. Further, method includes ligating a single-strand adapter with isolated complementary strand. Method is completed performing two successive PCR amplifications of ligated isolated complementary strand for determination of unknown polynucleotide sequence.

EFFECT: present invention improves sensitivity of detecting unknown DNA sequences, which flank known DNA sequence.

22 cl, 1 tbl, 7 ex