DNA Fingerprinting Forensic

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Presentation Transcript

DNA Fingerprinting Using PCR : 

DNA Fingerprinting Using PCR Timothy G. Standish, Ph. D.

Polymorphism : 

Polymorphism Differences between humans can be attributed to two factors: Environmental variation impacting development Individual genes that vary between people These genes which are variable are called polymorphic Each person is genetically unique because of their unique set of polymorphic genes All people are related in that the vast majority of their genes do not vary, but are identical from person to person

DNA Fingerprinting : 

DNA Fingerprinting DNA fingerprinting involves identification of DNA segments which vary between individuals A set of DNA fragments polymorphic enough to provide a unique set of fragments for all individuals, can be used to identify any specific individual in a population No single fragment will uniquely identify an individual, just as no single polymorphic genetic trait will uniquely identify a person, but a unique set of polymorphic DNA traits/fragments can serve as a reliable means of identification

Polymorphism In The TPA Gene : 

Polymorphism In The TPA Gene Tissue Plasminogen Activator (TPA) is a protein that functions in the cascade of reactions which break down blood clots The gene contains 14 exons and 13 introns Scattered within the introns are 28 Alu transposon sequences Within intron 8 a single Alu sequence may be present, or it may be missing Thus, as intron 8 varies with the presence or absence of Alu, it is polymorphic

Polymorphism In The TPA Gene : 

Polymorphism In The TPA Gene

PCR Detection of TPA Polymorphism : 

PCR Detection of TPA Polymorphism PCR will produce a 660 bp fragment 300 base pairs PCR will produce a 660 bp fragment if these primers are used

Components of a PCR Reaction : 

Components of a PCR Reaction Buffer (containing Mg++) Template DNA 2 Primers that flank the fragment of DNA to be amplified dNTPs Taq DNA Polymerase (or another thermally stable DNA polymerase)

PCR : 

PCR

PCR : 

PCR 3’ 5’ Heat

PCR : 

PCR

PCR : 

PCR Heat Heat 5’

PCR : 

PCR

PCR : 

PCR Heat Heat

PCR : 

PCR

PCR : 

PCR

DNA Between The Primers Doubles With Each Thermal Cycle : 

DNA Between The Primers Doubles With Each Thermal Cycle

PCR Detection of TPA Polymorphism : 

PCR Detection of TPA Polymorphism PCR will produce a 660 bp fragment 300 base pairs PCR will produce a 660 bp fragment if these primers are used

Gel Electrophoresis : 

Gel Electrophoresis