AMPLIFICATION OF NUCLEIC ACID LIBRARIES USING KINETIC EXCLUSION Russian patent published in 2021 - IPC C12Q1/6848 B01L3/00 

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to methods and a liquid system for increasing a number of amplicons in clonal populations of amplicons at amplification sites. A reaction of the first solution and the second solution differing from it is carried out on a flow cell in the liquid system by flowing the first solution through amplification sites located on the flow cell, and then by flowing the second solution through amplification sites. The first solution includes target nucleic acids and the first mixture of reagents, which includes nucleoside triphosphates and replication enzymes. Target nucleic acids in the first solution are transferred to amplification sites and bound to them at a transfer rate. Under the action of the first mixture of reagents, amplification occurs and amplicons are formed at an amplification rate that exceeds the transfer rate. The second solution includes the second mixture of reagents and does not contain any target nucleic acids. The second solution is intended for increasing the number of amplicons. Additional amplicons are formed by exclusion amplification at the amplification rate that exceeds the transfer rate.

EFFECT: inventions increase the number of amplicons in clonal populations or clusters at amplification sites, providing more complete formation of amplification clusters, and also provide a result with significantly higher intensity and higher signal/noise ratio, increasing the accuracy and/or efficiency of sequencing and analysis of the genetic library.

16 cl, 14 dwg, 3 ex