4C Polymerase Chain Reaction

• Denaturation: DNA heated to 90-95 degrees celsius to seperate strands
• Annealing: DNA cooled to 50-55 degrees celsius for primers to bind
• Elongation: DNA heated to 72 degrees celsius for Taq polymerase to create new strands
• The process of Polymerase Chain Reaction:

  1. Denaturation: at 90-95 degrees celsius
  2. Annealing: Cool down at 50-55 degrees celsius
  3. Elongation: at 72 degrees celsius
  4. Repeat
• Polymerase chain reaction requires a DNA sample, Taq polymerase, nucleotide bases, and sequence-specific DNA primers to function.
• Polymerase chain reaction is a DNA manipulation technique that amplifies DNA by making multiple identical copies, from a small initial sample. It involves thermal cycling. Polymerase chain reaction helps scientists run further analyses on the DNA.
• Forward primers: Bind to start codon at 3' end of the template strand, guiding synthesis in the same direction as RNA polymerase.
• Reverse primer: Bind to stop codon at 3' end of coding strand, guiding synthesis in the reverse direction to RNA polymerase.
• Having two primers is necessary because 5' end of template and coding strand differ. Also Taq polymerase functions only towards 3' end, so both primers are needed for directionality.