Helping Find the Missing Gene

Amanda Taylor and Jeffrey Johnson

June 13, 2003

About Our Talk 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 ExperimentFirst 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 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 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.

PCR is a method of testing the presence of DNA. A gel is used after the PCR to finish the detection.

Our Second Results 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.

1 3

Our Last Results 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 Radioactive Mouse Regular Mouse Groovy Mouse Campbell Mouse

Off To The Races

And the winner is, Groovy Mouse

Acknowledgements 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!