2. Helping Find the Missing Gene Amanda Taylor and Jeffrey Johnson June 13, 2003

3. About Our Talk The Purpose of Our Experiment Muscular Dystrophy The Mice Our Experiment Our Results A Special Surprise

4. 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.

5. 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.

6. The Mice Normal MDX Mouse

7. 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.

8. Making cDNA

9. 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.

10. 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.

11. 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

12. 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.

14. Mouse Race Radioactive Mouse Regular Mouse Groovy Mouse Campbell Mouse

15. Off To The Races And the winner is, Groovy Mouse

16. 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!