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GENE TECHNOLOGIES - Coggle Diagram

- - - - lab/ computerised machine
        
        identify a sequence -> work our amino acid sequence then mRNA and DNA sequence
        
        entered into the computer
        
        creates small sections of overlapping single strands which join to create DNA
      - V quick + accurate, can be intron free
    - - makes DNA copies from mRNA
        
        select a cell that naturally produces the protein of interest
        
        extracts mRNA
        
        reverse transcriptase (enzyme) joins DNA nucleotides with the complementary bases
        
        makes single stranded DNA (complementary DNA ) (cDNA)
        
        DNA polymerase is used to join free floating nucleotides to make double stranded DNA
      - V intron free
    - - cut up DNA at recognition sites
        -> naturally occur on bacteria
        
        creates sticky ends which are palindromic
        
        these ends can join DNA with complementary base pairs
  - - - isolate vector DNA
        
        cut open plasmids using same restriction endonucleases
        -> sticky ends are complementary
        
        join sticky ends together with ligase (ligation)
        -> catalyses condensation reaction
        
        insert into host cell (made more permeable)
        
        insert marker gene into vector (antibiotic resistance, fluorescence, enzymes)
      - if you want it too produce a specific protein - add promotor and terminator regions
    - - PCR - polymerase chain reaction
        
        in solution : DNA fragment, taq polymerase, primers, nucleotides
        
        95 c - break H bonds- single stranded DNA
        
        55 c - primers anneal
        
        72 c taq polymerase attaches to complementary nucleotides
  - - - AFTER RESTRICTION ENDONUCLEASES AND GEL ELECTROPHORESIS
        single stranded DNA is labelled radioactively or fluorescently
        patients DNA is heated to H bond break - single stranded
        binds with the probe if complementary
        can view under UV/ X ray
    - - DNA is heated to that it is a single strand
        mixed with complementary sequences of Single stranded DNA
        when cooled, they anneal
        used in medical diagnosis
    - - if sticks to dna probe for a mutation means the person has this mutation
      - useful for identifying inherited conditions
        
        huntingtons - symptoms not seen until over age of 30
      - useful for identification of health risks
        
        mutated alleles increase risk of developing cancer
  - - - found in introns
      - the more similar they are, the more realted the peope are