The Blotter from Down Under A l Represented by: Austin Walden and Tudor Hadade

Our History Founded by Edward M. Southern in 1975 Invented it while using radiolabeled DNA probe to detect a specific DNA sequence We also have sister companies Northern, Western, Eastern, and Southwestern blotting

What is Southern Blotting? Procedure used to identify specific DNA sequences Occurs after Gel Electrophoresis Utilizes Nucleic Acid Hybridization Uses a complementary DNA strand to find specific sequences and determine degree of hybridization

How We Operate Extract and purify DNA from cells Restrict DNA with restriction enzymes Separate into bands using Gel Electrophoresis Denature the DNA Transfer to a nitrocellulose paper Add labeled probe to allow for NA Hybridization Wash off unbound probe Observe using an Autoradiograph

Why We’re Important Through Nucleic Acid Hybridization, one may compare DNA and isolate target sequences Used in identifications of specific DNA Sequences Able to Identify and Name Mutations Identification for carriers of genetic disorders Prognosis of Cancer HIV Diagnosis DNA Fingerprinting

Cons Can be considered costly $250-500 per testing Reliant on a proper Gel Electrophoresis test Not Immediate Has a turnaround time of approximately 1-2 weeks

Pros Allows us to predetermine carriers for genetic disease We may then resolve and fix these disorders through other methods, such as gene therapy Effective in finding mutations in DNA Allows us to detect multiple genes in a genome Not strictly specific

Ethical Concerns Method itself has little debate over its practice After Southern Blotting has been performed, the information acquired from it may lead to some “questionable” experiments Gene therapy is a major usage and reason for debate Concerns over whether it is right or not to change a living organisms DNA Would it be right to change an unborn human’s DNA if we know they have a disorder? May make natural selection null for humans

Example Usage ETV6 disorder linked with defects of Short Arm part of Chromosome 12 (12p) Using Southern Blotting, it was discovered that: Rearrangement/Deletion of ETV6 gene occurs about 70% of the time in patients with an affected 12p 90% of children with a defected 12p/breakpoint in intron 5 were carriers for a rearrangement of ETV6 in intron 5 http://www.ncbi.nlm.nih.gov/pubmed/9665196