DNA Profiling Flashcards
Who first discovered DNA profiling?
Alex Jeffreys
What was the first use of DNA profiling?
Conviction of Colin Pitchfork, first mass DNA screening
What is DNA?
Hereditary material
Unique to each person (except identical twins)
Nucleotides- base, sugar, phosphate- C, A, G, T
Genetic code- provides information, which produces proteins
How is DNA organised?
DNA forms a gene, intervening sequences
Genes determine proteins, incorporated in chromosomes (stored in nucleus)
Mitochondrial DNA
What are possible sources of DNA?
Biological sources- saliva, hair root, cells, blood, semen, urine, tissues (i.e. skin, liver), bones, teeth
Trail of DNA- i.e. fingerprints, weapons, bitemarks
How much of a blood stain is required to obtain a DNA profile?
a very small amount
What is the method of RFLP to obtain a DNA profile?
Original method Restriction Fragment length polymorphism Variable number tandem repeats Large amounts of DNA required Requires under graded material (Warm moist conditions increase degradation)
What is the method of PCR to obtain a DNA profile?
Polymerase Chain Reaction
Short tandem repeats (1-5 bases, repeated 17 times)
Less DNA required
Can be partially degraded
Limitations- contamination from other sources, more sensitive
How does RFLP work?
Variable number tandem repeats
Patterns match
Restriction enzymes- cut at points of DNA
Separated through gel electrophoresis
How does PCR work?
DNA goes through a number of cycles, each cycle repeats the last one
Heated to high temperature- 95 degrees, causing DNA to separate
Lower temperate- introduce primers (primmer annealing/join) bind to specific parts of DNA
Increase temperature to 72 degrees- polymerase called Taq polymerase synthesises new DNA strand
Lots of copies of DNA- useful in degraded samples, can increase number of DNA
What is PCR used for?
To make lots of copies of specific sections of the DNA taken from biological samples
What are these specific sections known as?
STRs (Short Tandem Repeats)
What does one cycle of PCR go through?
Through a series of temperature changes
This cycle is repeated several times (usually 28) to produce more copies of the DNA
What is done once lots of copies of DNA are made?
Analyse it using gel electrophoresis
Primers labelled with fluroescent labels
Products exposed to laser beams
General facts about results from gel electrophoresis?
Less repeats of locus- smaller it’s going to be, faster travels through cell
Need to look at more than one locus
Thicker bands- same number repeats for each allele, allele from mother and father the same