Abstract: Provided are a highly specific Taq DNA polymerase variant and the use thereof in genome editing and gene mutation detection. All polar amino acids, directly interacting with a primer/template complex, on a Taq enzyme are selected to be mutated one by one to obtain 40 Taq variants, and extensive random mutagenesis is performed on the basis of the sequences of the variants and the sequences of wild type Taq enzymes to create a Taq mutant library. Then, a series of Taq mutants with a high specificity are screened on a qPCR screening system by means of taking a genome editing indels plasmid as a template, wherein the Taq mutants exhibit great advantages in CRISPR/Cas9 editing efficiency evaluation and single-cell cloning genotyping.
Type:
Application
Filed:
July 15, 2021
Publication date:
May 23, 2024
Applicants:
SHANGHAI TURTLE TECHNOLOGY CO., LTD., SHANDONG UNIVERSITY
Inventors:
Qilai HUANG, Xiaodan LIU, Ping DU, Bo LI