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Uses for Molecular Biology
Curing cancer, heart disease, Alzheimer's disease
Genetically manipulate corn for high yield & enhanced flavor
Genetically manipulate tomatoes for slower ripening
Identify criminals cases- murder or rape
enzymes that have been purified from a different species of bacteria and restriction enzyme
Cutting requires energy in form of ATP, involves physical cleaving of chemical bonds,
specific recognition, sites where cuts occur and are palodromic
Lab, Puc19 and E.Coli, relatively small extrocromisonal, circular molecule of DNA, found in bacteria and yeast
Puc19 used for cloning because of size and DNA sequence
Used to separate and sort fragments of DNA by size only, DNA fragments placed in gel, electric current runs through matrix, fragments move at different rates depending on protein's size and charge
Steps for cloning a gene and bacterial plasmid
1. Isolate DNA from two sources
2. Cut both DNA with some restriction enzyme
3. Mix DNA they join by base pairing
4. Add DNA ligase to bond covalently
5. Put plasmid into bacterium by transformation
6. Identify cells containing recombinant plasmid by ability to grow in presence of ampR and tetR
7. Clone selected bacteria
Used to stain gel after DNA samples were ran, allow visualization of discrete DNA bands on gel
recognize specific DNA sequence whenever it occurs in a DNA molecule and cut DNA near site, each restriction enzyme is named for the species of bacteria from which is located & isolated; Bacteria uses restriction enzymes to recognize & metabolize foreign DNA, assigned units based on amount required to digest 1 microgram of DNA in 1 hour
Molecular weight markers
Used to determine size of DNa fragments & Are of known size w/ which to compare results
buffer used for agarose gels electrophoresis in analysis of DNA products resulting from PCR amplification, using wrong concentration of TBE can result in gel and buffer over heating
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