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Transcript
Excruciating Extractions of Various Mouse Organs <Group A1> 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. www.promega.com . 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 Heart 180mg 18mg 275ml 250ml 650ml 20ug YES Bright Band Lungs(2) 200mg 20mg 275ml 250ml 650ml 20ug YES Faint Band Kidneys (2) 550mg 19mg 275ml 250ml 650ml 20ug YES Bright Smear Thymus 200mg 17mg 275ml 250ml 650ml 20ug YES Bright Smear Gel Electrophoresis of Purified DNA Heart Kidney Thymus Lung DNA Appearance in Agrose Gel Visibility 5 4 3 2 1 POSITIVE VALUES INDICATE STRENGTH OF BAND APPEARANCE 0 -1 -2 -3 -4 -5 NEGATIVE VALUES INDICATE STRENGTH OF SMEAR APPEARANCE Heart Kidneys Thymus Lungs 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.