1. Research Agenda  Examine mouse anatomy  Choose organ/tissue for structural/histological examination  Extract DNA from mouse tissue sample  Amplify DNA in PCR to identify a gene of interest (genotype)

2. Research Agenda  Run DNA Gel Electrophoresis to visualize genotyping results  Use bacterial transformation to express and replicate a gene of interest for in vitro studies  Check your transformation to verify gene presence using an enzyme digest assay  Examine protein localization in tissue using ImmunoHistoChemistry (IHC)

3. Mouse Anatomy  Secure mouse on styrofoam blocks with pins  Make a ventral midline incision, then cut across abdominal cavity, diaphragm and upper chest. You will need to cut through the sternum and ribs to expose heart and lungs

4. Mouse Anatomy  Locate the heart, lungs, diaphragm, liver, gal bladder, spleen, stomach, small intestines, cecum, large intestine, bladder, kidneys  Choose one organ to harvest for further histological examination: heart, brain, kidney, liver

5. Tissue Fixation  By fixation we intend to: (1) conserve the tissue from autolysis and bacterial attack (2) prevent the loss of cellular constituents (3) increase optical differentiation of cellular structures (4) increase tissue consistency, in order to facilitate their going through the other steps of the technique – especially slicing  How it works :  Paraformaldehyde will crosslink primary amino groups in proteins with other nearby nitrogen atoms in protein or DNA through a –CH 2 - linkage  A solution of 4% formaldehyde fixes pathology tissue specimens at about one mm per hour at room temperature  Depending on duration of exposure and temperature, fixation may be reversible or irreversible

6. Tail Snip  Cut a < 5 mm tail section and submerge it into DNA extraction buffer  Proteinase K enzyme buffer will digest tissue overnight and we will harvest the DNA tomorrow for further examination