THERMAL DISSOCIATION METHOD FOR SELECTION OF DNA-APTAMERS Russian patent published in 2020 - IPC G01N33/50 C12N15/115 

FIELD: biology.

SUBSTANCE: invention relates to molecular biology, enzymology, biotechnology and medicine and is intended for producing DNA-aptamers. Method includes: (a) pre-enrichment of the combinatorial DNA library by forming a complex thereof with an immobilized protein target (human alpha-fetoprotein; AFP) and conducting selection rounds; (b) forming a complex of the obtained library with a native (non-immobilized) AFP solution; (c) separating DNA / AFP complexes from unbound DNA oligonucleotides by acrylamide electrophoresis; (d) extracting AFP bound DNA from acrylamide gel at 37 °C; (e) thermal dissociation of the remaining AFP-complexed DNA from gel at 60 °C; (f) amplification of DNA after thermal dissociation. After the selection rounds with the use of thermal dissociation, a rich DNA library suitable for sequencing is obtained.

EFFECT: use of thermal dissociation, due to creation of more stringent washing conditions of DNA-oligonucleotides involved with the target, makes it possible to increase efficiency of selection of the most specific aptamers to protein targets.

3 cl, 4 dwg, 10 ex