6 results about "Taq DNA Polymerase" patented technology

This disclosure relates to a Taq polymerase-conjugated antibody composition and a method for nucleic acid amplification.

This invention belongs to the field of parasite detection technology, specifically relating to a lyophilized PCR kit for detecting Clonorchis sinensis in fecal samples. The lyophilized PCR kit of this invention includes a lyophilized reaction powder, a positive control, a negative control, and a reconstitution solvent. The lyophilized reaction powder is prepared by freeze-drying a reaction solution containing the following components: (a) a first primer pair and a first probe for detecting the Clonorchis sinensis COX1 gene; (b) a second primer pair and a second probe for detecting the human genomic actin (ACT) gene; (c) PCR reaction components: including hot-start Taq DNA polymerase, dNTPs, and a buffer system; and (d) a lyophilization protectant. This invention targets the Clonorchis sinensis-specific COX1 gene and uses PCR amplification technology to detect fecal samples, accurately identifying Clonorchis sinensis nucleic acid and effectively avoiding cross-reactions with other parasites and intestinal flora. The detection specificity and sensitivity are significantly improved, and the repeatability and stability are excellent.

The application belongs to the technical field of fluorescent quantitative PCR, and provides an enzyme digestion-fluorescent quantitative PCR detection method. The method comprises the following steps: step one, configuring a reaction mixture, wherein the reaction mixture comprises amplification reagents, a restriction endonuclease, Taq DNA polymerase, UDG enzyme, primers and a probe; and step two, setting a reaction program and a reaction system. Non-target templates in a qPCR reaction system are degraded by enzyme digestion of a restriction endonuclease, thereby reducing non-specific amplification; the proportion of target templates is increased, thereby avoiding the interference of non-target templates and improving detection sensitivity and detection stability. In particular, the method can effectively detect high GC content templates, such as TERT gene promoters or CpG island methylation.

The application discloses a Taq DNA polymerase mutant resistant to human whole blood and application thereof, relates to the field of biology, and the amino acid sequence of natural Taq DNA polymerase is shown as SEQ ID NO. 1. The mutant comprises all combination mutants of two or three of G395A, L408I and F413Y sites. The application is based on natural Taq DNA polymerase, and the molecular structure of the Taq DNA polymerase is reformed by rational design and a site-directed mutation biological technology, so that the Taq DNA polymerase mutant is more suitable for PCR amplification of samples containing human whole blood and the like than the wild type, and is more beneficial to the accuracy of result evaluation of samples containing human whole blood inhibitors.

A direct PCR detection method for trace colonies of Cryptococcus neoformans with gene mutations, belonging to the field of molecular biology detection technology, addresses the problems of existing technologies that require genomic DNA extraction, resulting in cumbersome steps and difficulty in handling trace samples. The method involves picking the Cryptococcus neoformans transformants to be tested, suspending them in PCR buffer, heating at 94°C for 12-15 minutes for thermal lysis, and then cooling on ice for 5-8 minutes for stability. Taq DNA polymerase, dNTPs, and primers are then directly added to the original PCR tube to prepare a complete PCR reaction system for amplification and electrophoresis detection. This invention eliminates the need for DNA extraction, purification, and sample transfer throughout the entire process, making it simple, fast, and cost-effective, especially suitable for single colonies or 10 colonies. 3 ~10 4 The experiment, which uses extremely small amounts of bacterial cells at the cell level, has a high success rate and can be completed in just 1-2 hours, making it highly valuable for high-throughput gene mutation screening and verification.