Amplifikationsverfahren und Reagenzsatz

Details

Veröffentlichungs-Nr.

EP4793363

Anmeldetag

12. Februar 2026

Veröffentlichung

19. August 2026

Rechtsraum

EP

IPC

C12Q1/6848

Abstract

The present application discloses a variety of amplification methods and reagent kits. In an amplification method provided in the present application, a first cycle of amplification is performed on a target polynucleotide through steps S20-S40, and an extended strand of a first amplification primer is generated on the surface of a solid carrier; then, amplification is performed on the extended strand of the first amplification primer through steps S60-S100, and an extended strand of a second amplification primer is generated on the surface of the solid carrier; subsequently, the extended strand of the first amplification primer and the extended strand of the second amplification primer are used as templates to perform cyclic amplification according to steps S120-S160 to obtain a molecular cluster of the target polynucleotides, so as to amplify signals generated by incorporating nucleotides or nucleotide analogs into the target polynucleotides in the process of sequencing the target polynucleotides, thereby making it easier to detect the signals and improving the efficiency and accuracy of signal detection. Meanwhile, by configuring at least one of betaine having a concentration greater than 0 mol/L and less than or equal to 4 mol/L, Mg<2+> having a concentration of 0.5-4 mmol/L, NH<4>+ having a concentration greater than 0 mmol/L and less than or equal to 100 mmol/L, and formamide having a percent concentration by volume (vol/vol% or v/v%) less than or equal to 15% in each of a first solution, a second solution, and a fourth solution, the amplification method provided in the present application can reduce the formation of a secondary structure in the template molecule, improve amplification efficiency, and facilitate the binding of the template molecule to the amplification primer and the extension of the amplification primer, thereby reducing GC bias.

Anmelder

Firma

GeneMind Biosciences Company Limited

Land

🇨🇳 China

Fachgebiete