Supercharge Your Innovation With Domain-Expert AI Agents!

Taq DNA polymerase mutant for probe-based qPCR

A polymerase and mutant technology, applied in the field of molecular biology, can solve the problems of limiting detection speed and separation, and achieve high efficiency

Active Publication Date: 2022-01-04
WUHAN AIBO TAIKE BIOTECH CO LTD
View PDF16 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, wild-type Taq DNA polymerase cannot dissociate the probe from its attached fluorophore within 1 second regardless of whether polymerization occurs during amplification, which limits detection speed

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Taq DNA polymerase mutant for probe-based qPCR
  • Taq DNA polymerase mutant for probe-based qPCR
  • Taq DNA polymerase mutant for probe-based qPCR

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0078] When using the Taq DNA polymerase mutant of the present invention for gene expression analysis, generally RNA is extracted from a sample, and then reverse-transcribed to obtain cDNA. Any method in the prior art can be used to detect the threshold of the reporter signal and determine the C of the sample. T , to achieve quantitative detection of cDNA targets.

[0079] get mutant

[0080] Taq DNA polymerase mutants were obtained by inverse PCR mutagenesis. All Taq DNA polymerase mutants in the present invention were expressed and purified in Escherichia coli, and were verified by sequencing, and for the convenience of purification, all mutants and wild-types were added with His tags at the C-terminus.

[0081] qPCR probe

[0082] Acquisition of qPCR probes: qPCR was performed with the SARS-CoN gene 2019-nCoV_N2 released by the 2019-nCoV real-time RT-PCR diagnostic team of the US Centers for Disease Control and Prevention as the target.

[0083] Forward primer: 2019...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a Taq DNA polymerase mutant for probe-based qPCR. In the present invention, Taq DNA polymerase mutants are designed, characterized and screened based on the probe method qPCR, and some Taq DNA polymerase mutants are obtained by adopting a cycle process of rapid amplification time (extension period of 1 second). Compared with wild-type Taq DNA polymerase, The mutants provided in the present application have the characteristics of rapid detection of amplification results, which significantly reduces the total time required for the qPCR process and improves the detection efficiency of qPCR or real-time qPCR. Therefore, the mutants provided by the present application have great economic advantages. The Taq DNA polymerase mutant provided by the present invention can be used for routine qPCR detection, including gene expression analysis and other DNA quantitative detection.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a Taq DNA polymerase mutant used for probe method qPCR. Background technique [0002] Taq DNA polymerase is generally used in the polymerase chain reaction (PCR) for the amplification of nucleic acid amplicons. In a PCR reaction, target DNA fragments (amplicons) are generally amplified through a cycle of three steps: denaturation, annealing, and extension. Real-time fluorescent quantitative PCR (qPCR) is to detect and record the amplification results by collecting the fluorescent signal data generated by dyes or probes during the PCR amplification process. Probe-based detection technology refers to the detection technology with the help of fluorescently labeled targeting probes. When the fluorescent probe is combined with the target sequence, the fluorescent dye is released, and the progress of the amplification can be inferred by real-time detection of changes in the ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12C12N15/54C12N15/63C12Q1/6851
CPCC12N9/1252C12Q1/6851C12Y207/07007C12N15/70C12R2001/19C12Q2561/113C12Q2563/107C12Q2531/113C12Q2521/107C12Q1/686Y02A50/30C12Q2521/101C12Q1/6853
Inventor 朱振宇孙大鹏
Owner WUHAN AIBO TAIKE BIOTECH CO LTD
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com