Welcome to Sindh Biotechnologist Association

Polymerase Chain Reaction (PCR) is one of the top-most innovation and technique in the field of biotechnology and has aided to the success of different biotechnological and medical research and tests. The PCR technique, developed by Kary Mullis in 1980s, permits scientists to make multiple copies of a target DNA region, simplifying the analysis, and the manipulation of genetic material. A heat-resistant enzyme called DNA polymerase is used for replicating DNA in cycles of heating and cooling. Polymerase chain reaction is an amplification process through which a small amount of nucleic acid (DNA or RNA) can be multiplied and further tests can be performed. The machine in which the process is performed is termed as thermo-cycler and the process of PCR was first introduced by Kary Mullis, a renowned American Biochemist   (Plantsurfer, 2023) . Principles and working of PCR: Polymerase Chain Reaction (PCR) takes place in a series of 3 steps i.e.: Denaturation, Annealing, and El...

Background Using a DNA template, at least one pair of unique primers, deoxyribonucleotides, an effective buffer solution and a thermostable DNA polymerase, the same technique is used to amplify small quantities of DNA as in traditional PCRs. In a thermal cycler containing sensors for measuring the fluorescence of the fluorophore after excitation at the appropriate wavelength, a material marked with a fluorophore is applied to this mixture, enabling the generation rate to be measured for one or more particular items. This makes it possible to calculate the generation rate of the amplified product during each PCR period. The data thus created can be analysed in several samples by computer software to measure relative gene expression (or copy number of the mRNA). In order to assess the occurrence and abundance of a given DNA sequence in these samples, quantitative PCR can also be extended to the identification and quantification of DNA in samples. This calculation is made after each am...