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Biology LibreTexts

8.S: Techniques of Molecular Genetics (Summary)

  • 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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