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Genetic Engineering (Reaction Endonucleases (Nucleases break down…

- - - - Identify insulin cDNA using probes
        
        Add sticky ends to cDNA (GGG)
        
        Preparation of a vector
        
        Remove plasmid with antibiotic resistance gene and promoter + terminator from bacteria by dissolving bacteria cell wall then centrifuging and extracting plasmid
        
        Cut up plasmid using restriction endonuclease
        
        Add sticky ends (CCC)
        
        1 more item...
  - - - Remove mRNA from cell expressing the gene
        
        mRNA + reverse transcriptase = single stranded cDNA
        
        cDNA + DNA polymerase = double stranded cDNA
    - - Use amino acid sequence to determine the DNA base sequence
        
        Computer builds oligonucleotides which are added together to bulld the gene
        
        Gene copied by PCR and given sticky ends
- - - - A sterile velvet pad, the same size as the petri dish, is then pressed onto the dish, picking up a sample of each colony
        
        The bacteria can then be stamped onto new plates, in the identical arrangement
        
        The medium in the plates contains antibiotics - either one or two different ones
        
        So colonies can be identified that are antibiotic resistant - therefore the desired bacteria can be isolated