You are on page 1of 1

Human DJ-l was modified by oxidation, specifically at residue, C106; and the ability of DJ-1 to be oxidized was a prerequisite

for the antioxidant properties of the protein. However, the extent of additional DJ-1 residues was unclear that may have acted to modulate DJ-1 oxidation. This research performed a comprehensive sitedirected mutagenesis study, which altered DJ-1 amino acid residues that hypothesized to be oxidized in cells. The total number of oxidation-sensitive amino acid residues in DJ-1 was predicted. Each group chose an amino acid residue in the DJ-1 sequence to mutate the oxidation sensitive residue into an oxidationinsensitive residue, M26. The section of the research focused on the creation of primer designs. However, too much substances added to the M26 primer would produce many cultures. Thus, it is likely by diluting the primer concentration would inhibit the making of the PCR product. The M26 primer was set up and ran PCR (polymerase chain reaction), DpnI digested- a restriction enzyme which digested methylated DNA- of PCR product, resolved on agarose gel confirming the existence of the product, transformed, inoculate mini cultures, minipreped the DNA, and the DNA was quantified. It was discovered that compared to other primers, assumed that M26 resulted in many cultures, nevertheless, it was not the cultures visible but contamination that happened throughout the research. Furthermore, the engagement of adding more than the required substances to designing the primer not only caused dilution but contamination along the way of the research. Taken together, these findings suggest that although M26 did not perform the correct comprehensive site-directed study, another issue conveyed what would result in the dilution of the primer.