PCR Gel ElectrophoresisIntroductionSickle cell anemia is caused by a single nucleotide polymorphism (SNP). SNPs are the most common type of genetic variation, as each SNP represents a difference in one nucleotide. SNPs are normally found throughout a person's DNA. More commonly, they are usually found in non-coding regions and have no effect on health. When a SNP occurs in a coding region, it can play a direct role in disease by influencing gene function. This sickle cell anemia SNP produces the recognition sequence for a restriction enzyme, HindIII. Samples without SNPs (non-disease carriers) do not contain the recognition sequence and are not cut. This allows the digested DNA fragments to be analyzed by separating them with the use of gel electrophoresis, allowing the possibility of identifying whether a patient suffers from sickle cell disease. Before we can visualize DNA, we must amplify the DNA. To amplify the DNA we will use a technique called polymerase chain reaction (PCR). With the invention of the PCR technique, DNA profiling has made enormous strides in both discriminatory power and the ability to recover information from very small (or degraded) initial samples. PCR greatly amplifies quantities of a specific region of DNA, using primers and DNA polymerase. The PCR method is easily adaptable to analyze STR loci. The PCR method relies on repeated ED heating and cooling cycles of the reaction for DNA denaturation and enzymatic replication of DNA primers containing complementary DNA in the target region, with DNA polymerase synthesis the reaction. The DNA polymerase used is normally Taq polymerase as it is very thermostable. As the PCR progressed, the DNA generated… half the paper… with no bands, proving that there was no contamination. These results imply that the experiment was a success as no problems were found. All five samples were used to increase the reliability of the experiment. DNA-free PCR is used as a negative control, there was no band expected to ensure there is no effect when there should be no effect. Uncut DNA PCRs should be used to compare cut and uncut DNA. It also ensures that there is an effect when there should be. An improvement to the experiment could include a digest containing fragments of known sizes to use as a guide to confirm that the cleaved DNA is the expected size. References http: //peds.oxfordjournals.org/content/13/4/283.full http://ghr.nlm.nih.gov/handbook/genomicresearch/snphttp://www.ndsu.edu/pubweb/~mcclean /plsc431/ students/firas.htm