MindMap Gallery Principles and applications of PCR technology
Molecular Biology, Human Health 9th Edition, PCR (Polymerase Chain Reaction): It is a molecular biology technology used to amplify specific DNA fragments. It can be regarded as special DNA replication outside the organism.
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Avatar 3 centers on the Sully family, showcasing the internal rift caused by the sacrifice of their eldest son, and their alliance with other tribes on Pandora against the external conflict of the Ashbringers, who adhere to the philosophy of fire and are allied with humans. It explores the grand themes of family, faith, and survival.
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[Zootopia Character Relationship Chart] The idealistic rabbit police officer Judy and the cynical fox conman Nick form a charmingly contrasting duo, rising from street hustlers to become Zootopia police officers!
Principles and applications of PCR technology
concept
PCR (Polymerase Chain Reaction): It is a molecular biology technology used to amplify specific DNA fragments. It can be regarded as special DNA replication outside the organism.
Features
Highly sensitive
High specificity
high yield
Repeatable
Quick and easy
Low requirements for samples
working principle
PCR system
Template DNA
Specific primers (upstream and downstream primers)
dNTP
Thermostable DNA polymerase (such as taq DNase)
Contains mg² buffer
distilled water
Basic reaction steps of PCR
①Transsexuality
The reaction system is heated to 95°C to completely denature the template DNA into a single strand, and at the same time, the primers themselves and the local double strands between the primers are also eliminated.
②Annealing
Lower the temperature to a suitable temperature (generally 5°C lower than Tm) to allow the primers to bind to the template DNA.
③Extension
The temperature is raised to 72°C, and DNA polymerase uses dNTP as a substrate to catalyze the DNA synthesis reaction.
PCR product analysis method
①Electrophoresis analysis
②Enzyme digestion analysis
③Hybridization analysis
④Sequence analysis
⑤ High performance liquid chromatography analysis
Main uses of PCR technology
① Obtain the target gene fragment
②Micro analysis of DNA and RNA
③DNA sequence analysis
④ Gene mutation analysis
⑤In vitro mutation of genes
Commonly used PCR techniques
①Reverse transcription PCR technology
②In situ PCR technology
③Quantitative PCR (real-time PCR technology)