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PCR Flow Chart
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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.