Applications in research and diagnostics

• What is the primary purpose of PCR?

  To make millions of copies of DNA
• What is PCR used for in research?

  To make lots of copies of genes
• How can contamination affect PCR results?

  It can lead to inaccurate amplification
• What is the purpose of the annealing step in the PCR process?

  • Annealing (55°C) allows the DNA primers to bind to the complementary sequences on the template DNA.
• What is the function of DNA polymerase in PCR?

  It builds new DNA strands
• What is a key requirement for PCR to work effectively?

  Specific primers
• What is the purpose of DNA denaturation in the PCR process?

  • DNA denaturation (95°C) separates the double-stranded DNA into single strands to allow primers to bind.
• What is the role of DNA polymerase in the PCR process?

  • DNA polymerase synthesizes new DNA strands complementary to the template DNA strand.
• How long does it take to perform PCR?

  Just a few hours
• What is the temperature used for the annealing step in the PCR process?

  55°C
• What happens during the denaturation step of PCR?

  Heat separates the DNA into single strands
• What role do primers play in PCR?

  They attach to the target DNA strands
• How does the PCR cycle contribute to DNA amplification?

  It is repeated to exponentially amplify DNA
• What does PCR stand for?

  Polymerase Chain Reaction
• What is a use of PCR in DNA sequencing?

  Amplifying DNA for accurate sequencing
• What contamination risk is associated with PCR?

  Can lead to false positives if not careful
• How does PCR assist in diagnosing conditions?

  By amplifying DNA/RNA to detect pathogens
• What are the advantages and limitations of using PCR?

  Advantages:

  • High sensitivity: Detects minimal DNA
  • Rapid amplification: Millions of copies in hours
  • Cost-effective: Reduces need for large samples
  • Versatile: Applications in sequencing and engineering

  
  

  Limitations:

  • Requires specific primers: Need target sequence
  • Contamination risk: False positives possible
  • DNA polymerase errors: Inaccuracies in DNA
  • Limited amplicon size: Short DNA fragments only
• What are the four types of nucleotides (dNTPs) used in the PCR process?

  dATP, dCTP, dGTP, dTTP
• What does PCR do to DNA from a single copy?

  Amplifies it to billions in hours
• What are the basic steps of the PCR process?

  1. Denaturation: Heat separates DNA into strands.
  2. Annealing: Primers attach to target DNA.
  3. Elongation: DNA polymerase builds new strands.
• What is the name of the enzyme used in the PCR process?

  Taq
• What are the main applications of PCR?

  • Research Applications:
  • Genetic Engineering
  • DNA Sequencing
  • Gene Function Studies
  • Diagnostic Applications:
  • COVID-19 Testing
  • Genetic Disorder Detection
  • Cancer Diagnosis
• Why are DNA primers used in the PCR process?

  They provide a starting point for DNA synthesis by DNA polymerase
• What is the result of PCR cycles?

  Amplification of DNA from a single copy
• How does PCR help scientists study genes?

  By amplifying genes for analysis
• How is PCR used in genetic engineering?

  By modifying specific DNA sequences for research
• What is the temperature used for the denaturation step in the PCR process?

  95°C
• If a researcher wants to study a specific gene's function, what can they use PCR for?

  To amplify that gene for experiments
• What are the limitations of PCR?

  Requires specific primers and is contamination-sensitive
• What is one research application of PCR?

  Genetic Engineering
• What is a key advantage of PCR regarding sensitivity?

  It detects minimal amounts of DNA
• What is a limitation of PCR regarding amplicon size?

  Can only amplify relatively short DNA fragments
• What is the purpose of the repeat cycle (20-40 times) in the PCR process?

  To exponentially amplify the target DNA sequence
• Why is PCR important for detecting disease-causing organisms?

  It makes trace amounts detectable
• How does PCR work in COVID-19 testing?

  It amplifies viral RNA from nasal swabs
• What conditions can PCR help diagnose?

  Infections, genetic disorders, and cancer
• What are some advantages of using PCR?

  Its speed and accuracy
• What is the significance of amplifying DNA in PCR?

  It allows detection of low quantities of pathogens
• What issue can DNA polymerase errors cause in PCR?

  Can introduce inaccuracies in the amplified DNA