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8.S: Techniques of Molecular Genetics (Summary)

  • Page ID
    4115
  • [ "article:topic", "authorname:tnickle" ]

    • Molecular biology involves the isolation and analysis of DNA and other macromolecules
    • Isolation of total genomic DNA involves separating DNA from protein and other cellular components, for example by ethanol precipitation of DNA.
    • PCR can be used as part of a sensitive method to detect the presence of a particular DNA sequence
    • PCR can also be used as part of a method to isolate and prepare large quantities of a particular DNA sequence
    • Restriction enzymes are natural endonucleases used in molecular biology to cut DNA sequences at specific sites.
    • DNA fragments with compatible ends can be joined together through ligation.If the ligation produces a sequence not found in nature, the molecule is said to be recombinant.
    • Transformation is the introduction of DNA (usually recombinant plasmids) into bacteria.
    • Cloning of genes in E. coli is a common technique in molecular biology, since it allows large quantities of a DNA for gene to made, which allows further analysis or manipulation
    • Cloning can also be used to produce useful proteins, such as insulin, in microbes.
    • Southern blotting can be used to detect the presence of any sequence that matches a probe, within a mixture of DNA (such as total genomic DNA).
    • The stringency of hybridization in blotting and in PCR is dependent on physical parameters such as temperature and washing solution content.

    Key Terms:

    macromolecules

    lysis

    detergent

    chelating agent

    EDTA

    nuclease

    pellet

    PCR

    primer

    thermalcycle

    denature

    anneal

    extend

    Taq DNApol

    electrophoretic gel

    restriction endonuclease

    EcoRI

    sticky end

    blunt end

    compatible end

     

    ligation

    ligase

    plasmid

    transformation

    competent

    electroporation

    selectable marker

    agarose

    vector

    band

    ethidium bromide

    Southern blot

    membrane

    denaturation

    hybridization

    washing

    probe

    stringency

    northern blot

    western blot

    transgene

    GMO

    transformation

    transfection

    naked DNA

    vector

    electroporation

    microinjection

    vesicles

    callus

    knock-out

    recalcitrant

    Agrobacterium

    particle bombardment

    T-DNA

    Ti plasmid

    position effects

    stem cells

     

     

     

     

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