Manipulating genomes

    Cards (34)

    • What is the purpose of PCR?
      To amplify fragments of DNA
    • Why is PCR important for crime scene analysis?
      It amplifies insufficient DNA samples
    • What components are needed to prepare a PCR mixture?
      • DNA sample
      • Free DNA nucleotides
      • Primers
      • DNA polymerase from thermophilic bacteria
    • At what temperature does DNA separation occur in PCR?
      95°C
    • What happens during the annealing stage of PCR?
      The primer binds to the DNA
    • What is the optimum temperature for DNA polymerase during PCR?
      72°C
    • How does DNA polymerase contribute to DNA synthesis in PCR?
      It catalyzes phosphodiester bond formation
    • How many cycles of PCR are typically carried out?
      30-40 cycles
    • What is the role of ethidium bromide in electrophoresis?
      It binds to DNA and makes it visible
    • Why does DNA move towards the positive charge in gel electrophoresis?
      DNA is negatively charged
    • How do shorter DNA fragments behave in gel electrophoresis?
      They travel further than larger fragments
    • What are the steps involved in gel electrophoresis?
      1. Prepare agarose gel with wells
      2. Place gel in buffer solution
      3. Mix DNA with loading dye
      4. Pipette DNA into wells
      5. Apply electrical current
      6. Visualize banding pattern under UV light
    • What do restriction enzymes do?
      Cut double-stranded DNA at specific sequences
    • What are palindromic recognition sites in DNA?
      They read the same forwards and backwards
    • What are the two ways restriction enzymes can cut DNA?
      Creating blunt ends or sticky ends
    • How do scientists use restriction enzymes in genetic manipulation?
      To extract genes from longer DNA sections
    • What is DNA profiling used for?
      To analyze and compare DNA samples
    • What is the first step in DNA profiling?
      Collecting a DNA sample
    • How are PCR products separated in DNA profiling?
      Using gel electrophoresis
    • What can DNA profiling reveal about individuals?
      Genetic relationships and evolutionary links
    • What is the process of genetic engineering for insulin production?
      1. Remove insulin gene from human DNA
      2. Cut plasmid with restriction enzymes
      3. Join sticky ends with DNA ligase
      4. Mix recombinant plasmid with bacteria
      5. Use electroporator to increase permeability
      6. Grow transgenic bacteria in fermenters
    • How are plants genetically modified to produce drugs?
      • Create GM bacterium with drug gene
      • Bacterium infects plant cell
      • Inserts DNA into plant genome
      • Plant grows with drug-producing gene
      • Drug can be extracted or consumed
    • How are genetically modified animals produced?
      • Inject gene into fertilized egg cell
      • Implant into adult animal
      • All cells develop with drug-producing gene
      • Protein is purified from milk
    • What are the arguments for and against GMOs?
      Arguments for:
      • Increased crop yield and nutrition
      • Pest-resistance reduces pesticide use
      • Human proteins reduce allergic reactions
      • Vaccines stable without refrigeration
      • Cheaper industrial processes

      Arguments against:
      • Patents make seeds expensive
      • Risk of creating 'superweeds'
      • Ethical concerns about manipulation
      • Religious objections to 'playing God'
      • Unforeseen long-term impacts
    • What is gene therapy used for?
      To treat genetic diseases
    • How is gene therapy applied for recessive diseases?
      A functional gene copy is introduced
    • What is the difference between germ line and somatic gene therapy?
      Germ line affects gametes; somatic affects body cells
    • What are the problems associated with somatic gene therapy?
      Requires multiple insertions and may trigger immune response
    • What is the chain-termination method used for?
      To sequence DNA
    • What are the steps in the chain-termination method for DNA sequencing?
      1. Mix DNA sample, primers, polymerase, nucleotides
      2. Add modified nucleotides with fluorescent tags
      3. Carry out PCR to create varying lengths
      4. Use electrophoresis to separate fragments
      5. Read sequence from gel
    • What is the purpose of high-throughput sequencing?
      To sequence DNA quickly and cheaply
    • What is whole genome sequencing?
      • Fragment DNA using restriction enzymes
      • Insert fragments into bacterial artificial chromosomes (BACs)
      • Insert BACs into bacteria for replication
      • Purify and further fragment DNA
      • Sequence shorter fragments and assemble genome
    • What is synthetic biology?
      • Use DNA sequences to build proteins
      • Example: Synthetic artemisinin from yeast
    • What are the applications of comparing DNA sequences?
      1. Epidemiology: Identify disease-linked genes
      2. Evolutionary relationships: Determine organism closeness
      3. Genotype-phenotype relationships: Predict traits from genotype
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