公开(公告)号：US20250084447A1

公开(公告)日：2025-03-13

申请号：US18649214

申请日：2024-04-29

Applicant: EAST CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY

Inventor： Shenlin Wang ,  Mengbing Zou ,  Changxing Ma

IPC: C12P19/34 ,  C12N15/70

Abstract: Provided is a method for preparing stable isotope-labeled single-stranded DNA (ssDNA) by biosynthesis with E. coli, and the 15NH4Cl or 13C-Glcose is used as the only nitrogen or carbon source, which may significantly reduce costs. In the method of the present disclosure, the target sequence of ssDNA is tandemly duplicated on a high-copy vector, a site for a first restriction endonuclease and a site for a second restriction endonuclease are added to the 5′ and 3′ ends of the target sequence, respectively, and the recombinant vector is digested to obtain an asymmetric double-stranded DNA structure, which is then isolated by denaturation to obtain two ssDNAs of unequal lengths, including 15N- or 13C-labeled target ssDNA. The method of the present disclosure is able to effectively increase the yield of ssDNA, thereby improving the efficiency of in vitro synthesis of isotope labeled ssDNA.