Manipulating Genomes

    Cards (33)

    • What is PCR used for?
      To amplify fragments of DNA
    • Why is PCR important for crime scene analysis?
      It amplifies insufficient DNA samples
    • What components are needed to prepare for PCR?
      • DNA sample
      • Free DNA nucleotides
      • Primers
      • DNA polymerase from thermophilic bacteria
    • At what temperature does DNA strands separate 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 in PCR?
      72°C
    • How does DNA polymerase function during PCR?
      It catalyzes phosphodiester bond formation
    • How many cycles of PCR are typically carried out?
      30-40 cycles
    • What is the purpose of adding a fluorescent molecule to DNA?
      To visualize DNA under UV light
    • How does gel electrophoresis separate DNA fragments?
      By length, due to DNA's negative charge
    • What are the steps 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?
      Sequences that read the same forwards and backwards
    • What are the two ways restriction enzymes can cut DNA?
      Blunt ends or sticky ends
    • How do scientists extract a gene of interest using restriction enzymes?
      By analyzing recognition sites around the gene
    • 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 determine besides criminal identification?
      Genetic relationships between individuals
    • What is the process of making insulin using genetic engineering?
      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. 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 synthesizes the drug
    • How are genetically modified animals produced?
      • Inject gene into fertilized egg nucleus
      • Implant into adult animal
      • All cells develop with drug gene
    • What are the arguments for using GMOs?
      • Increase crop yield and nutrition
      • Reduce pesticide use
      • Produce human proteins without allergies
      • Stable vaccines for remote areas
      • Cheaper industrial enzymes
      • Cost-effective drug production
    • What are the arguments against using GMOs?
      • Patented seeds are 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
    • What is the difference between germ line and somatic gene therapy?
      Germ line affects gametes; somatic affects body cells
    • What are some problems with gene therapy?
      Immune response, insertion errors, and cost
    • What is the chain-termination method used for?
      To sequence DNA
    • How does the chain-termination method work?
      1. Mix DNA sample, primers, polymerase, nucleotides
      2. Add fluorescently-labelled modified nucleotides
      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)
      • Amplify BACs in bacteria
      • Purify and sequence fragments
      • Use software to reconstruct genome
    • What is synthetic biology?
      • Building proteins from DNA sequences
      • Example: Synthetic artemisinin from yeast
    • What are the applications of comparing DNA sequences?
      1. Epidemiology: Identify disease-linked genes
      2. Evolutionary relationships: Assess organism relatedness
      3. Genotype-phenotype relationships: Predict traits
    See similar decks