Excruciating Extractions of Various Mouse Organs Meghan Ellwanger, Brandi Golden Lisa Malmgren, Nathan Pineault, Amanda Steffens

Disclaimer The mouse used for this experiment was euthanized in full accordance to USDA and ICAUC ethical standards. The Wizard SV Genomic DNA Purification System is a registered trademark of the Promega Corp. Purpose Technique Results Data Summary Conclusion Q and A

Introduction Over the past few weeks we used the Promega “Wizard” kit to purify samples of extracted DNA obtained from various mouse organs The DNA was tagged with Ethidium bromide to illustrate the difference in DNA concentrations between organs. Gel electrophoresis allowed for visualization of DNA from the varying organ tissues.

Purpose To analyze the differences in DNA assay in different organ tissues of the body To familiarize and gain practical knowledge of common laboratory technique To successfully organize and interpret the data that is collected during the laboratory practical work.

Procedure 1.Extract organs from mouse 2.Digestive solution was used to disrupt tissues and release genomic DNA 3.After incubation, Lysis buffer, containing chaotropic salt, was added to the samples These salts denature cellular proteins and faccilitate the binding of the nucleic acids to the silica membrane. 4.The individual samples were washed repeatedly in the wash solution. 5.Eluted DNA was viewed using gel electrophoresis. 6.The compiled data was used to analyze the different organs’ DNA

Organs & Additives Mouse Organs Whole Weight Amount of Tissue used Digestive Solution Lysis buffer Wash Sol. DNA + 4ml Dye DNA Obtained Band Appearance Heart180mg18mg275ml250ml650ml20ug YESBright Band Lungs(2)200mg20mg275ml250ml650ml20ug YESFaint Band Kidneys (2) 550mg19mg275ml250ml650ml20ug YESBright Smear Thymus200mg17mg275ml250ml650ml20ug YESBright Smear

Gel Electrophoresis of Purified DNA HeartKidneyThymusLung

DNA Appearance in Agrose Gel POSITIVE VALUES INDICATE STRENGTH OF BAND APPEARANCE NEGATIVE VALUES INDICATE STRENGTH OF SMEAR APPEARANCE

Conclusion We were able to successfully obtain visual representation of DNA from all extracted organ samples. The Hypothesis is that the appearance of DNA on the gel correlates to the multiplicity and complexity of functions within the various organs. The brightest, most prominent band/smear indicates a higher concentration of DNA.