Scientists around the world have wondered for many years what distinguishes speciation. Of particular interest is the genetic basis for human/primate (chimpanzee or gorilla) separation. Humans and chimpanzees are 99% identical in their genomic DNA sequence, thus making them very closely related. Despite this high degree of sequence similarity, humans and primates have a number of striking phenotypic differences. We hypothesize that sequence changes that have occurred between humans and primates have altered developmental programs. Because transcription factors alter the expression of numerous genes, we also hypothesize that changes in the expression or activity of transcription factors are responsible for the different phenotypic traits among humans and primates. Using human chromosome 22 as a model for comparison between human and primate DNA, a random selection of noncoding genes approximately 1-2 kilobases (kb) long upstream was sequenced. Focused on promoter regions from the sequence data, significant differences were detected when comparing humans and gorillas (p-value= < 0.01) and gorillas and chimpanzees (p-value= < 0.01) suggesting that limited similarities existed between the species. When comparing humans and chimpanzees (p-value= > 0.1), no significant difference was detected. Using this information, transcription factors were analyzed between the human and chimpanzee data to determine if transcription regulation was different between the species. The results indicated no significant difference between humans and chimpanzees at the single-nucleotide level even though the species differ at the genetic and phenotypic levels. The results also indicated that changes in transcription regulation have played a major role in determining speciation. This research opens new avenues in investigating how many of the differences have functional consequences and the relative contributions of these transcription factors to expression differences.