In-vitro cloning (PCR)

• What does PCR stand for?

  Polymerase Chain Reaction
• What is the purpose of PCR?

  To amplify DNA fragments in vitro
• What does "in vitro" mean?

  Outside of a living organism
• What is the role of the thermocycler in PCR?

  It automates the temperature changes for PCR
• What is the first step in the PCR process?

  Denaturing the DNA at 95 degrees C
• What happens during the denaturing step of PCR?

  Hydrogen bonds between DNA strands break
• What temperature is used to allow primers to align?

  55 degrees C
• What is the purpose of primers in PCR?

  To bind to complementary DNA sequences
• What enzyme is used in PCR?

  DNA polymerase
• Why is Taq polymerase used in PCR?

  It withstands high temperatures without denaturing
• What is the optimum temperature for Taq polymerase?

  72 degrees C
• What happens during the synthesis stage of PCR?

  DNA polymerase attaches nucleotides to form DNA
• How many copies of DNA are produced after one cycle of PCR?

  Two copies of the DNA fragment
• What are the key stages of the PCR process?

  1. Denaturing: DNA strands separate at 95°C
  2. Annealing: Primers bind at 55°C
  3. Synthesis: DNA polymerase adds nucleotides
• What is a significant advantage of using PCR?

  It rapidly produces billions of DNA copies
• In what field is PCR also used besides recombinant DNA technology?

  Forensic science for DNA analysis
• Why is PCR considered less complex than in vivo cloning?

  It does not require living cells
• What are the advantages of using PCR in genetic analysis?

  • Rapid amplification of DNA
  • Requires no living cells
  • Useful for small DNA samples
  • Automated process increases efficiency