Optimizing the condition of the PCR detecting of PTEN gene in DNA Research Paper - 1

Optimizing the condition of the PCR detecting of PTEN gene in DNA derived from human cell lines, associated with ageing and senescence - Research Paper Example Traditional PCR is used for amplifying DNA for sequencing, genotyping and cloning. However, this has a number of shortcomings namely; poor precision, low sensitivity, short, dynamic range of less than 2 logs, low resolution, is non-automated, based on discriminatory size only, results cannot be expressed as numbers, the staining agent ethdium bromide is not efficient at quantification and finally, post-PCR processing is required. This paper looks at the analysis of PTEN using traditional PCR method. The process was not successful despite the necessary steps being undertaken. PTEN primer, when used on a Hacat cell line, showed annealing at approximately 62 degrees. However, the use of PTEN with similar parameters that were later altered for optimization yielded no results. The methodology has been discussed, and the gel results showing synthesized images have been included in this report. The discussion that covers the PCR process and its effectiveness has also been included. The fidelity of the PCR process has been discussed to extrapolate on the possible reasons for the failure of this analysis. Phosphatase and Tensin Homolog gene commonly referred to as PTEN gene provides the necessary instructions for the synthesis of a protein that is present in almost all body tissues (Sulis, 2003). The protein is significant in its function as a tumor suppressor and aids in the regulation of cell division by inhibiting cells from diving and growing too fast or uncontrollably. PTEN is a phosphotase which modifies other fats and proteins through the removal of phosphate groups comprising of one phosphorus atom and three oxygen atoms (Cohen 1990). There are a number of medical conditions that are linked to PTEN gene mutation including : Cowden Syndrome and Bannayan-Riley-Ruvalcaba syndrome(Weary et al, 1972; Cohen 1990) which are often termed together as the PTEN hematoma tumor syndrome. The gene mutations that are responsible for these conditions are found in body