Polymerase Chain Reaction

Polymerase Chain Reaction - high temperatures will be used...

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Polymerase Chain Reaction (PCR) The polymerase chain reaction can be used to make many copies of small pieces of DNA. Because techniques in biotechnology usually require large amounts of DNA (many copies), PCR has allowed much of the biotechnology development that we have seen in recent years. Materials and Procedure Materials needed The procedure requires primers, DNA polymerase, and nucleotides. Primers are short chains of about 20 nucleotides that are complimentary to a region in the DNA to be amplified. They are needed because the enzyme that copies the DNA (DNA polymerase) cannot start the process unless it has already been started. DNA polymerase from the thermophile Thermus aquaticus is used because this species thrives at temperatures that are near boiling. It's DNA polymerase (called Taq polymerase) is stable at relatively high temperatures and functions optimally at 70 degrees C. This is important because
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Unformatted text preview: high temperatures will be used to separate the strands of the double helix. Nucleotides are needed because DNA is composed of nucleotide "building blocks". Procedure The DNA in question is heated to approximately 95 degrees C to separate the two strands of the double helix. After the strands are separated, the DNA is cooled to about 50 degrees and the primers attach. The temperature is raised to approximately 70 degrees C. This is the optimal temperature for Taq polymerase. The polymerase attaches to and copies the strand. The solution is then heated and cooled again as described above at regular intervals. Each time it is cycled through this heating and cooling procedure, the DNA replication process repeats itself and the amount of DNA produced is doubled....
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This note was uploaded on 01/04/2012 for the course BSC BSC1005 taught by Professor Orlando,rebecca during the Fall '10 term at Broward College.

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Polymerase Chain Reaction - high temperatures will be used...

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