What is the thermal cycling process in PCR?

Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. It can be considered a special form of DNA replication outside of a living organism. The main feature of PCR is its ability to significantly increase trace amounts of DNA.

Overview of a Polymerase Chain Reaction Cycle

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What is the thermal cycling process in PCR?

PCR consists of three basic steps:

1. Denaturation of Template DNA: The template DNA is heated to around 93°C for a certain duration, causing the double-stranded DNA to separate into single strands, preparing it for primer binding in the next reaction cycle.

2. Annealing of Template DNA and Primers: After denaturation, the temperature is lowered to around 55°C, allowing the primers to bind to the complementary sequences on the single-stranded template DNA.

3. Extension of Primers: At 72°C, the DNA template-primer complex undergoes extension with the help of DNA polymerase (like Taq DNA polymerase), using dNTPs as substrates to synthesize a new complementary strand based on the template DNA.

Beijing LIUYI PCR Machine

By repeating the denaturation, annealing, and extension processes, more “semi-conservative replication strands” are generated, which can serve as templates for subsequent cycles. Each cycle takes about 2 to 4 minutes, allowing the target gene to be amplified millions of times within 2 to 3 hours.

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