Download DNA Analysis of Various Mouse Organs

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.