Polymerase Chain Reaction (PCR)
Polymerase Chain Reaction (PCR) has become one of the most versatile and widely used techniques in Molecular Biology. By combining specific primers and probes, genes or segments of nucleic acids (DNA/RNA) can be amplified for downstream applications in as little as an hour. Because the PCR reaction results in an exponential replication of DNA, as little as a few copies of the gene of interest can be replicated into billions of copies by the end of the reaction. These copies can be quantified against known standards analyzed concurrently to determine the starting amount of the gene of interest.
Since initial development of the method, the PCR process has been adapted and modified into numerous new methods including Real-time PCR (rt-PCR), Quantitative PCR (qPCR), Reverse transcriptase PCR (RT-PCR), Multi-plex PCR and Nested PCR, among others. EAG utilizes rt-PCR for the determination of genetic sex in the Japanese Medaka and the African clawed frog as well as quantitative RT-PCR to detect levels of Vtg1 in the Japanese Medaka.