Dan Ririe, Department of Chemistry

Introduction

Within the mouse genus Peromyscus there exists many pericentric inversions containing linkage groups, genes which do not recombine. It has been hypothesized that genes inherited together provide the selective pressure which has caused speciation within the genus. The purpose of this project was to aid ongoing research in the lab of Dr. Duke Rogers in finding genetic markers in the mouse genus Peromyscus. From these markers a linkage map will eventually be developed, It is this linkage map which can help answer several evolutionary questions concerning this genus.

Methods

I used a technique requiring small I 0-base primers of random sequence to amplify products from mouse DNA with polymerase chain reaction (PCR). This technique is known as RAPDS, randomly amplified polymorphic DNA, and has been used in many projects to successfully create a linkage map in relatively short amount of time.

Using DNA samples collected by other researchers I screened twenty RAPD primers for polymorphism. In my screening I tested two protocols. The protocol I used to collect data was an adaptation by one printed by Scott Woodward (1991). An alternative procedure used originally on plant DNA studies also showed readable markers.

Results and Conclusion

The data was analyzed using the Chi-square statistics test and resulted in five usable markers. The most significant results from this project came from the improved method of producing RAPD markers. The original protocol used was improved and an alternative procedure was attempted. Although not the intent of this study, the experimentation with the protocol for plant DNA revealed valuable information; by using primers of a certain GC content (greater than 70%), this procedure works well with mammalian DNA. This procedure will be useful in the continued work that is needed to complete this project.