DNA Fingerprinting Techniques

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Presentation transcript:

DNA Fingerprinting Techniques

RFLP Stands for Restriction Fragment Length Polymorphism Takes advantage of differences in DNA between individuals that result in different size fragments when digested with restriction enzymes* Cut specific DNA sequences How many fragments will result when each of these alleles are digested with DdeI? 3 fragments 2 fragments

RFLP To see RFLP, DNA is digested with the appropriate restriction enzymes and run on an agarose gel in a process called electrophoresis.

Gel Electrophoresis Agarose gels are porous and will allow the DNA fragments to move through them Electric current (running between a negative to positive pole) applied to the gel will cause the DNA (slightly negative charge) to be pulled through the gel towards the positive pole DNA fragments will be separated by size

Which fragments will move the fastest? Small or Large? Smaller fragments will move faster and farther down the gel than larger fragments Once separated by electrophoresis, DNA fragments can be visualized using various techniques

Ethidium Bromide Staining EtBr is a chemical that binds to DNA and glows in UV light. We can “stain” our DNA bands by soaking the gel in EtBr. When we expose the gel to UV light we will see a distinct banding pattern – a DNA fingerprint.

RFLP Animations Animation #1 Animation #2

Time to Practice “DNA Goes to the Races”

Interpreting a Paternity Test DNA fingerprint Each individual inherits 50% of each parent’s DNA. Half of your RFLP’s came from mom and half from dad We can compare the DNA fingerprints of parents and children to determine paternity

What can we learn from the cases below? Mom Baby Dad Mom Baby Dad Are mom and dad homozygous or heterozygous? Is baby homozygous or heterozygous? Could mom and dad be the parents? Could mom and dad be the parents? YES YES

Activity Time Let’s “Find a Family” using DNA fingerprints