Patent Document ID: 9862991
Application ID: 14384960
Patent Flag: 1

Claim One:
1. A method of gene synthesis, comprising the steps of: (1) immobilizing a pool of oligonucleotides to the surface of a solid substrate to form a gene chip, wherein the oligonucleotides are anchored to the surface of the chip via a linker sequence that is 15-150 bases in length, located at the 3′ end, and contains a nicking endonuclease recognition site; (2) carrying out oligonucleotide pool amplification on the gene chip by performing an isothermal nicking and strand displacement amplification reaction, which comprises performing the following steps under isothermal conditions: (i) hybridizing a universal primer to the linker at the 3′ end of the chip-immobilized oligonucleotides, (ii) extending the universal primer using a strand displacement polymerase to form a newly amplified oligonucleotide chain, (iii) nicking the newly amplified oligonucleotide chain using a nicking endonuclease to re-free the 3′ end of the universal primer for a new extension reaction, and (iv) using the strand displacement polymerase to extend the universal primer of step (iii) and displace the newly amplified strand generated in step (ii), and (3) carrying out parallel gene assembly on the gene chip by performing a polymerase cycling assembly reaction using the newly amplified oligonucleotide chains generated in the oligonucleotide pool amplification step.