公开(公告)号：US20230228696A1

公开(公告)日：2023-07-20

申请号：US17951411

申请日：2022-09-23

Applicant: EAST CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY

Inventor： Shenlin Wang ,  Sha Zhao

IPC: G01N24/08

CPC classification number: G01N24/087

Abstract: An experimental technology for detecting a hydrogen bond based on an ssNMR technology includes: (1) exciting a 1H nucleus of an RNA sample with a π/2 pulse; (2) applying two π pulses every half rotation period on an X_nucleus of the RNA sample; (3) applying a π pulse on the 1H nucleus of the RNA sample; (4) applying two π pulses every half rotation period on the X nucleus of the RNA sample; (5) applying a 90° pulse on 1H and X atoms of the RNA sample; (6) recording a chemical shift of X in indirect dimension; (7) applying the 90° pulse on the 1H and X nuclei of the RNA sample; (8) repeating steps 2, 3, and 4; and (9) collecting the 1H signal in direct dimension; where X is selected from the group consisting of 15N and 13C.

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.

公开(公告)号：US11971376B2

公开(公告)日：2024-04-30

申请号：US17951411

申请日：2022-09-23

Applicant: EAST CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY

Inventor： Shenlin Wang ,  Sha Zhao

IPC: G01N24/08

CPC classification number: G01N24/087

Abstract: An experimental technology for detecting a hydrogen bond based on an ssNMR technology includes: (1) exciting a 1H nucleus of an RNA sample with a π/2 pulse; (2) applying two π pulses every half rotation period on an X nucleus of the RNA sample; (3) applying a π pulse on the 1H nucleus of the RNA sample; (4) applying two π pulses every half rotation period on the X nucleus of the RNA sample; (5) applying a 90° pulse on 1H and X atoms of the RNA sample; r a chemical shift of X in indirect dimension; (7) applying the 90° pulse on the 1H and X nuclei of the RNA sample; (8) repeating steps 2, 3, and 4; and (9) collecting the 1H signal in direct dimension; where X is selected from the group consisting of 15N and 13C.