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PCR Flow Chart

PCR Flow Chart

Kevin Alonso

In the Laboratory

The most commonly used apparatus in the labs to amplify segments of DNA via the polymerase chain reaction is the PCR machine also known as the thermal cycler

Denature DNA

Samples are heated to 94-96 degree Celsius for one to several minute to denature (separate into single strands) the targets DNA

Anneal Primers

The temperature is lowered to 50-56 degree Celsius for one to several minutes. This allows the DNA primers to anneal(base pair) to their complementary sequences. The primers are designed to bracket the DNA region to be applied.

Extend Primers

The temperature is raised to 72 degree Celsius for one to several minutes. This allows Taq polymerase to attach at each priming site and extend a new DNA strand completing two double-stranded copies of the target DNA

Copies; Repeat

The PCR reaction mixture contains many copies of the primers and an abundant supply of DNA nucleotides/dNTPs to perform many addition cycles. Do these steps again about 30+ Times. Each cycle takes about 2 hours to do and the more cycles the better.

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