Watson & Crick

James Watson & Francis Crick (1953) Using data from Chargaff & Franklin, they determined that DNA must be a double (not single) helical polymer

Modeling DNA
Through construction of a model, Watson & Crick determined that by using the complimentary base pairing of A-T & C-G, DNA could attain a double helix shape

The Significance of Watson & Crick

Provided an explanation for how all the parts of DNA could be arranged in a manner that would allow the molecule to replicate itself 1st Part of Central Dogma

The Complimentary Base Pairing Of The Nitrogenous Bases Is Due To The Number of Hydrogen Bonds

During prophase, the supercoils of DNA may be further condensed into chromosomes

How Does DNA Copy Itself?

REPLICATION OF DNA

 DNA molecule uncoils from tightly condensed state, assisted by DNA Helicase. This unzipping occurs at the replication fork. Hydrogen bonds between nitrogenous bases break New exposed half-rungs of ladder-like structures are called TEMPLATES Free DNA nucleotides in the nucleus are brought into place by DNA POLYMERASE
Due to complimentary base pairing, only free Adenines can pair with Thymines in the template, only free Guanines can pair with Cytosines in the template, etc.

REPLICATION

DNA Polymerase also proofreads the strands for mispaired nucleotides. The DNA fork is recoiled

Why Is It Essential That The Hydrogen Bonds Between Nitrogenous Bases Be Weak?

Replication Occurs In Two Directions Simultaneously

Lab: Extraction of DNA

Purpose: To ascertain and appreciate several properties of DNA based upon the processes required to extract DNA from the nucleus of a eukaryotic cell. Methodology: Proteolytic enzyme extraction and separation using non-polar solvent

Wheat Germ
Source of Nucleic Acids Wheat germ is derived from the part of the wheat seed that is undergoing high mitotic activity

Lauryl Sulfate
Active ingredient in dishsoaps Why necessary to disrupt the plasma and nuclear membranes?

Heat
Why is it important to have heat? Why is it important not to exceed 50C?

Papain & Bromelin

Proteolytic enzymes found in meat tenderizer. Derived from bromeliad plants (i.e. pineapple) that use these enzymes to ward off parasites Problem: Meat tenderizer also contains nucleases

Sodium Hydrogen Carbonate

NaHCO3 Disassociated sodium ions bind to negative aspects of DNA polymer, protecting it from nucleases in meat tenderizer Can produce carbonic acid when it disassociates. Why might this be useful in terms of papain?

Ethanol
Ethanol is non-polar. DNA is water soluble Why use ethanol to extract DNA?

Lab Protocol
Add 45mL tap water to a 100 mL test tube and place in a hot water bath. Heat until the temperature of the water INSIDE THE TEST TUBE is 55C. DO NOT, AT ANY POINT, EXCEED 60C! Add 2g wheat germ and 3mL of lauryl sulfate (detergent) to the 45mL of water in the test tube. Stir and incubate for 5 minutes

Lab Protocol 2
After the first incubation, add 2g of Papain (meat tenderizer) and 5mL 1M NaHCO3. Stir and incubate at 55C for 15 minutes. NEW STEP: Filter the liquid portion of the mixture through a coffee filter into a 100 mL beaker. Cool the beaker to 25 C in an ice bath When ready, instructor will layer in 20mL iced ethanol. Let stand 3-5 minutes and observe.