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Lecture 15

Lecture 15 – Key Developments in DNA Post RFLP.docx

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Anthropology 2235A/B
Eldon Molto

Lecture 15 – Key Developments in DNA Post RFLP  RFLP was time consuming and required considerable amounts of DNA to be used  1984 - New technology the Polymerase Chain Reaction (PCR - the DNA Xerox machine) allowed for smaller amounts of DNA (usually found at crime scenes) to be amplified thereby preserving the sample  In 1991 Edwards writes first paper on new technique called Short Tandem Repeats, which unlike RFLP could be adapted to the new PCR technology!  Single Nucleotide Polymorphisms  Note on Taq Polymerase o Taq polymerase is a heat tolerant enzyme that is derived from the thermophilic bacteruium Thermus aquaticus first isolated from a oceanic thermal vent by Thomas Brock in the 1980s! This enzyme must be heat tolerant because heat-fragile enzymes would have to be replaced after every step. Our enzymes only operant at body temperature  PCR Reaction o To multiply a DNA molecule with PCR requires:  Target DNA molecule  Primers  DNA polymerase that directs replication  Nucleotides (4 bases) o Steps involved in PCR  High heat to separate the double helix (denaturing)  Cooling to allow primer binding (anneling)  Slight warming to allow polymerase to bind bases to the target (extension)  STR Nomenclature o If a marker falls within a gene (exon) the gene name is used e.g. the STR TH01 is from the human tyrosine hydroxylase gene located on chromosome 11, the ’01’ portion of this STR comes from the fact that the repeat region in question is located within intron (non-coding) 1 of the
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