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Helping Find the Missing Gene
Amanda Taylor and Jeffrey Johnson
June 13, 2003
About Our Talk
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The Purpose of Our Experiment
Muscular Dystrophy
The Mice
Our Experiment
Our Results
A Special Surprise
The Purpose of Our Experiment
The purpose of our experiment was to
make cDNA libraries from a healthy mouse
and a mouse with muscular dystrophy. A
cDNA library is a collection of all the
active genes in a tissue. These two libraries
can be used to study muscular dystrophy by
helping find genes involved with the
disease.
What is Muscular Dystrophy?
Muscular Dystrophy is a genetic disorder
described by muscle wasting and weakness
that begin with very small changes in the
muscles. As a person’s muscles die over
time, the person’s muscle strength lessens,
eventually stopping the heart and causing
the person to die.
The Mice
Normal
MDX Mouse
Our Experiment
First we took the RNA from two mice, a
healthy one and a diseased one. We then
proceeded to use the RNA to make cDNA, an
exact copy of the RNA except that it is in
DNA form. Afterwards we embarked on the
journey of cleaning the cDNA and inserting it
into bacteria.
Making cDNA
Our First Results
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Our first attempt
failed. We came in
and we found no
bacteria growing.
This meant that none
of the cells had taken
up the DNA so they
could grow. We had
to start over.
Our Second Attempt
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We made the cDNA
over, being more
careful not to lose our
samples again. We
then used PCR to test
the cDNA that we
made to make sure
that there was
actually some present.
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PCR is a method of
testing the presence
of DNA. A gel is used
after the PCR to
finish the detection.
Our Second Results
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1
3
This is a picture of a gel we ran to
see the DNA made in the PCR.
Lane 1 shows the DNA made by
PCR from the normal mouse and
lane 3 shows no DNA was made
from the diseased mouse.
Since we only had supplies to try
making one more library, we used
the normal mouse cDNA to finish
our experiment.
Our Last Results
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We purified the cDNA and used PCR to
see which sample from the purification
process would be the best to use to
complete the library.
When we tried to put the purified cDNA
into bacteria, the bacteria did not grow. So
no library, again.
Mouse Race
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Radioactive Mouse
Regular Mouse
Groovy Mouse
Campbell Mouse
Off To The Races
And the winner is, Groovy Mouse
Acknowledgements
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Dr. Bloom, Pharmaceutical Sciences Dept.
Mrs. Bloom, STEP Organizer
My partner
Campbell University
Everyone else that helped us with our
project! Thank you very much!