Invitro Cloning

• Polymerase Chain Reaction


  Uses an automated machine (thermocyler), DNA fragments, DNA polymerase (taq), primers and nucleotides

  • Temperature increased to 95C, to break hydrogen bonds and split DNA to single strand
  • Temperature decreased to 55C so primers can anneal
  • Temperature increased to 72C for optimum, DNA polymerase (taq) attaches complementary free nucleotides (synthesis)
• Advantages of PCR

  • Automated - more efficient
  • Rapid - 100 billion copies within hours
  • No living cells - quicker and less complex