LIFESCI 3 Chapter Notes - Chapter 7.2: Polymerase Chain Reaction, Reverse Transcriptase, Real-Time Polymerase Chain Reaction
Document Summary
Gene sequences can be amplified with the polymerase chain reaction. Polymerase chain reaction (pcr): amplifies the number of copies of a dna segment. Can be used if we know the end portions of the sequence we are interested in. Dna polymerases synthesize dna strands on a pre-existing dna template using free deoxyribonucleotides. Two oligonucleotides complementary to sequences on opposite strands of the target. Serve as replication primers that can be extended by a dna polymerase. 3" ends of hybridized primers are oriented toward each other and positioned to prime. Dna sample containing the segment to be amplified. Rxn mixture is heated to separate dna strands cooled so primers anneal to dna segment is replicated by dna polymerase using dntps. Repeated 25 - 30 times to amplify dna segment until it can be analyzed/cloned. Exponential growth (# segments doubles every time) Dna polymerase is heat stable and remains active after each step.
