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Exam (Genes in Medicine – linkage analysis (What is polymerase Chain…

- - - - Hold large fragments of DNA
      - Acts to increase the number of copies through replication
      - Plasmid (Grow in bacteria and only small pieces of DNA)
        
        pUC19
        
        Features
        
        an ampR selectable marker
        
        Multiple cloning site
        
        located within part of the B-Galactosidase gene lacZ
        
        Contain multiple different restriction palindromes into which DNA fragments can be inserted/cloned
        
        Origin of Replication (Ori)
        
        lacZ gene acts as an indicator of successful gene insertion as...
        
        Bacterial colonies with pUC19 grown on nutrient agar with lactose present will either be...
        
        Blue which indicates unsuccessful insertion as the colonies have fermented lactose meaning lacZ is still intact
        
        White which indicates successful insertion as the colonies have not fermented lactose.
        
        Recombinant DNA technology
        
        Restriction enzyme used to make sticky ends on the Plasmid so that gene of interest can be inserted
  - - - Contains DNA fragments representing the entire genome of an organism or species.
      - Process
        
        DNA is cut into cloneable sized pieces as randomly as possible using restriction endonucleases
        
        Cloning resulting fragments into a vector makes a gDNA Library which contains
        
        Introns
        
        Promoters
        
        Origins of replication
        
        Exons
      - Screening
        
        Colony Hybridisation
        
        Using
        
        Radioactivety or Antibiotics
    - - Contains Coding Sections only
      - Synthesisation
        
        Synthesised by using messenger RNA as a template
        
        synthesised during reverse transcription
  - - - They are single base-pair variations
      - • Most of human diversity is in the form of SNPs
      - • In humans, SNPs occur about once in 100-300 bases
      - • 90% of human genome variation comes from SNPs