PCR TESTING
Nona Scientific Molecular
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FASTER TESTING
What is Polymerase Chain Reaction (PCR)
Polymerase Chain Reaction (PCR) is a method to analyze a short sequence of DNA (or RNA) even in samples containing only minute quantities of DNA or RNA. PCR is used to reproduce (amplify) selected sections of DNA or RNA. Previously, amplification of DNA involved cloning the segments of interest into vectors for expression in bacteria and took weeks. But now, with PCR done in test tubes, it takes only a few hours. PCR is highly efficient in that untold numbers of copies can be made of the DNA. Moreover, PCR uses the same molecules that nature uses for copying DNA:
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Primers
Two "primers", short single-stranded DNA sequences that are synthesized to correspond to the beginning and ending of the DNA stretch to be copied
Polymerase
An enzyme called polymerase that moves along the segment of DNA, reading its code and assembling a copy
DNA Blocks
A pile of DNA building blocks that the polymerase needs to make that copy
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VISUAL SIMPLICITY
How is PCR Done?
There are three major steps involved in the testing process (view animated picture of a PCR). Each step is repeated for 30 or 40 cycles. The cycles are done on an automated cycler, a device that rapidly heats and cools the test tubes containing the reaction mixture. With one cycle, a single segment of double-stranded DNA template is amplified into two separate pieces of double-stranded DNA. These two pieces are then available for amplification in the next cycle. As the cycles are repeated, more and more copies are generated, and the number of copies of the template is increased exponentially. Each step – denaturation (alteration of structure), annealing (joining), and extension – occurs at a different temperature:
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DOWNLOADABLE RESOURCES
Nona Molecular PCR handouts
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