A Real-Time Quantitative PCR Assay for Quantification of c-Myc DNA in Patients who Suffers from Leukemia

Abstract

 The MYC cancer gene contains instructions for the production of the c-Myc protein. The c-Myc protein is known as a transcription factor or a regulator of other genes. It is a protein that binds DNA at specific sites and instructs genes whether or not they should be transcribed into messages for cells to make additional or other new proteins. Quantitative real-time PCR (qRT-PCR) addresses the evident requirement for quantitative data analysis in molecular medicine, biotechnology, microbiology, archaeometry and diagnostics and has become the method of choice for the quantification of cDNA and nDNA. Therefore, we used Polymerase chain reaction (PCR)-based assays can target either DNA (the genome) or cDNA, namely used for research both DNA. We optimized a method for monitoring quantitative real-time PCR (qRT-PCR) of c-Myc cancer gene in patients with leukemia. We describe qRT-PCR a series of protocols that illustrate the essential technical steps required to generate quantitative data that are reliable and reproducible. In addition, our aim is to also quantify extracted DNA and determine its purity and the validation of extracted DNA from patients with leukemia including active Myc gene family. We also believe these protocols will be accessible to the researchers to provide them reliable data in this protocol. These analytical methods are essential for accurate gene quantification. With reference to, advantages of qRT-PCR are a large dynamic range of quantification, no requirement for post-PCR sample handling and the need for very small amounts of starting material. The specificity, reproducibility and detection limit of the assay was examined. The assay was used to monitor c-myc DNA levels in patients with leukemia.